Apc^Q8, Apc2^g10 homozygous follicle cell clones lead to an increase in the numbers of polar/stalk cells, as compared to controls.

ApepP^EY02585 homozygous or heterozygous progeny from ApepP^EY02585 heterozygous parents do not reach adulthood when also homozygous for Apc2^g10, inherited from heterozygous parents.

Apc^Q8/Apc^Q8, Apc2^g10/Apc2^g10 mutant intestinal stem cell clones in the adult posterior midgut are significantly larger, with more cells, than control wild type clones of similar age, and form multi-layered structures bulging in the lumen of the gut. This phenotype is visible 10 days after clone induction, but becomes more prominent with time. As compared to controls, there is an increase in the number of apoptotic cells in close proximity to the Apc^Q8/Apc^Q8, Apc2^g10/Apc2^g10 clones, affecting enterocytes, enteroendocrine cells, and intestinal stem cells. Wild type clones are dramatically smaller and fewer in number in the presence of Apc^Q8/Apc^Q8, Apc2^g10/Apc2^g10 clones, as compared to wild type clones in control guts.

Expression of Diap1^Scer\UAS.cHa under the control of Scer\GAL4^bun-GSG5961 and Scer\GAL4^GSG5966 (along with RU486 to induce expression via the GeneSwitch system) in the gut fully restores growth of wild type clones in the presence of Apc^Q8/Apc^Q8, Apc2^g10/Apc2^g10 mutant clones in the posterior midgut, and suppresses the growth of Apc^Q8/Apc^Q8, Apc2^g10/Apc2^g10 mutant clones; however, Apc^Q8/Apc^Q8, Apc2^g10/Apc2^g10 mutant clones expressing Diap1^Scer\UAS.cHa under the control of Scer\GAL4^tub.PU do not have suppressed growth.

Expression of puc^Scer\UAS.cMa or bsk^DN.Scer\UAS under the control of Scer\GAL4^bun-GSG5961 and Scer\GAL4^GSG5966 (along with RU486 to induce expression via the GeneSwitch system) in the gut fully restores growth of wild type clones in the presence of Apc^Q8/Apc^Q8, Apc2^g10/Apc2^g10 mutant clones in the posterior midgut, and suppresses the growth of Apc^Q8/Apc^Q8, Apc2^g10/Apc2^g10 mutant clones. Apc^Q8/Apc^Q8, Apc2^g10/Apc2^g10 mutant clones expressing puc^Scer\UAS.cMa or bsk^DN.Scer\UAS under the control of Scer\GAL4^tub.PU also show suppressed growth. Expression of puc^Scer\UAS.cMa or bsk^DN.Scer\UAS in host tissue only, under the control of Scer\GAL4^GSG2326, severely suppresses growth of Apc^Q8/Apc^Q8, Apc2^g10/Apc2^g10 mutant clones.

Expression of Myc^Scer\UAS.cZa in host midgut tissue only, under the control of Scer\GAL4^GSG2326, fails to suppress the overgrowth of Apc^Q8/Apc^Q8, Apc2^g10/Apc2^g10 mutant clones or the reduced growth of wild type clones.

hpo^42-47/+ or ex^e1/+, but not yki^B5/+, fully suppresses the reduced growth seen in posterior midgut wild type clones when in the presence of Apc^Q8/Apc^Q8, Apc2^g10/Apc2^g10 mutant clones, and suppresses the growth of Apc^Q8/Apc^Q8, Apc2^g10/Apc2^g10 mutant clones.

Apc^Q8/Apc^Q8, Apc2^g10/Apc2^g10 mutant clones expressing yki^{Scer\UAS.T:Avic\GFP-EGFP,T:Ivir\HA1} under the control of Scer\GAL4^tub.PU do not have suppressed growth.

Cultured primary neurons derived from doubly homozygous Apc2^g10/Apc2^g10 ; Apc^Q8/Apc^Q8 embryos have a significantly increased axon length compared to wild-type controls. This phenotype is unaffected if the embryos are also homozygous for CLIP-190^KO.

Animals maternally and zygotically mutant for both Apc^Q8 and Apc2^g10 are embryonic lethal, the embryos are smaller and display denticle loss while the naked cuticle is expanded.

Apc^Q8 Apc2^g10 double maternal and zygotic mutant embryos exhibit a severe cuticle phenotype characterized by a complete loss of denticles, anterior head holes caused by failure of head involution, and an overall reduction in size caused by increased apoptosis.

Expression of Apc2^{WT.T:Avic\GFP-EGFP} suppresses the embryonic lethality and cuticle patterning defects seen in maternally and zygotically Apc2^g10 Apc^Q8 double mutant embryos.

Expression of Apc2^{ΔR1R3-R5.T:Avic\GFP-EGFP} largely suppresses the embryonic lethality and cuticle patterning defects seen in maternally and zygotically Apc2^g10 Apc^Q8 double mutant embryos.

Expression of Apc2^{Δ15Δ20R1R3-5.T:Avic\GFP-EGFP} does not suppress the embryonic lethality and only modestly improves the cuticle patterning defects seen in maternally and zygotically Apc2^g10 Apc^Q8 double mutant embryos.

Expression of Apc2^{Δ15Δ20R1R4-5.T:Avic\GFP-EGFP} substantially suppresses the embryonic lethality and cuticle patterning defects seen in maternally and zygotically Apc2^g10 Apc^Q8 double mutant embryos.

Apc2^g10 Apc^Q8 mutant intestinal stem cell clones are significantly larger in size than wild type controls. On day 14 after clone induction the clones begin to develop into multilayered epithelia, and by day 21 many clones have fused with each other, outcompeting the wild type cells to occupy the majority of the midgut. Clone cell polarity appears normal.

Expression of pan^ΔN.Scer\UAS under the control of Scer\GAL4^Act.PU suppresses the increased size of Apc2^g10 Apc^Q8 mutant intestinal stem cell clones 7 days after clone induction, suppressing the development to hyperplasia. Multilayered clones are rarely observed, even on day 21.

Expression of pygo^KK108114 under the control of Scer\GAL4^Act.PU suppresses the increased size of Apc2^g10 Apc^Q8 mutant intestinal stem cell clones 14 days after clone induction.

Expression of sgg^Scer\UAS.cUa under the control of Scer\GAL4^Act.PU suppresses the increased size of Apc2^g10 Apc^Q8 mutant intestinal stem cell clones 14 days after clone induction, preventing hyperplasia.

Expression of Egfr^DN.Scer\UAS under the control of Scer\GAL4^Act.PU suppresses the increased size of Apc2^g10 Apc^Q8 mutant intestinal stem cell clones 7 and 14 days after clone induction (ACI). The multilayering phenotype often seen at 21 days ACI is also suppressed.

Ras85D^e1B suppresses the increased size of Apc2^g10 Apc^Q8 mutant intestinal stem cell clones 7 and 14 days after clone induction (ACI). The multilayering phenotype often seen at 21 days ACI is also suppressed. This reduction in cell number is due to reduced proliferation, rather than increased cell death: TUNEL expression is unchanged and expression of the apoptotic protein BacA\p35 has no effect on the phenotype. The remaining increase in clone size seen in the Ras85D^e1B, Apc2^g10, Apc^Q8 triple mutant flies is fully suppressed upon expression of pan^ΔN.Scer\UAS under the control of Scer\GAL4^Act.PU, and no increase in apoptosis is seen in these flies.

Flies co-expressing Ras85D^V12.Scer\UAS produce fewer intestinal stem cell (ISC) clones per midgut over time compared with Apc2^g10 Apc^Q8 mutants alone, similar to what is seen when Ras85D^V12.Scer\UAS is expressed alone. Approximately 35% of the clones that remain undergo rapid proliferation and develop into large spherical-tumor cell masses (termed 'transformed clones'). There is no obvious increase in apoptosis in these transformed clones and the majority are found in the anterior or posterior midgut rather than the middle midgut. Few enterocytes (ECs) or enteroendocrine cells (ees) are seen, indicating that differentiation is blocked. The remaining 65% of clones are smaller than their wild type counterparts, but the ratio of ISCs to differentiated EC and ee cells is comparable to wild type.

Approximately 10% of Apc2^g10 Apc^Q8, Ras85D^V12.Scer\UAS mutant ISC clones extrude basally toward the surrounding muscle layer, although the underlying base layer remains unbroken. Expression of Ras85D^V12.Scer\UAS induces polarity changes in Apc2^g10 Apc^Q8 mutant clones.

Expression of phl^{Scer\UAS.F179} induces cell polarity changes in Apc2^g10 Apc^Q8 mutant intestinal stem cell clones.

Expression of hep^Act.Scer\UAS under the control of Scer\GAL4^Act.PU suppresses the hyperplasia seen in Apc2^g10 Apc^Q8 mutant intestinal stem cell clones.

Expression of shg^NIG.3722R under the control of Scer\GAL4^Act.PU enhances the formation of multicellular epithelia seen in Apc2^g10 Apc^Q8 double mutant clones. Enterocytes and enteroendocrine cells are both present and the ratio of differentiated cells to progenitor cells is similar to that observed in Apc2^g10 Apc^Q8 clones alone. The apicobasal polarity of the clone cells is also unaffected.

23% of embryos derived from females carrying homozygous Apc2^g10 Apc^Q8 germline clones and heterozygous double mutant males (generated using the FRT-FLP-DFS technique) are able to hatch. All of the surviving embryos are maternally double null and zygotically heterozygous for the double-null chromosome. A further 25% of embryos display weak cuticle defects and these are also zygotically heterozygous for Apc2^g10 and Apc^Q8. All of the maternally and zygotically double mutant embryos die showing severe cuticle defects.

Expression of Apc2^{FL.T:Avic\GFP-EGFP} partially suppresses the lethality seen in embryos derived from females carrying homozygous Apc2^g10 Apc^Q8 germline clones and heterozygous double mutant males (generated using the FRT-FLP-DFS technique). 46% of embryos are able to hatch. The remaining progeny exhibit a very weak cuticle phenotype.

Expression of Apc2^{Δ20RΔB.T:Avic\GFP-EGFP} fails to suppresses the lethality seen in embryos derived from females carrying homozygous Apc2^g10 Apc^Q8 germline clones and heterozygous double mutant males (generated using the FRT-FLP-DFS technique). 24% of embryos are able to hatch and the severity of the cuticle defects in the remaining progeny is similar to the double mutant alone.

Expression of Apc2^{Δ15RΔ20RΔB.T:Avic\GFP-EGFP} fails to suppress the lethality seen in embryos derived from females carrying homozygous Apc2^g10 Apc^Q8 germline clones and heterozygous double mutant males (generated using the FRT-FLP-DFS technique). Between 18% and 24% of embryos are able to hatch, depending on the expression level of the transgenes. The severity of the cuticle defects in the remaining progeny is similar to the double mutant alone.

Expression of Apc2^{R3-R5SA.T:Avic\GFP-EGFP} partially suppresses the lethality seen in embryos derived from females carrying homozygous Apc2^g10 Apc^Q8 germline clones and heterozygous double mutant males (generated using the FRT-FLP-DFS technique). 68% of embryos are able to hatch.

Expression of Apc2^{R3-R5SD.T:Avic\GFP-EGFP} partially suppresses the lethality seen in embryos derived from females carrying homozygous Apc2^g10 Apc^Q8 germline clones and heterozygous double mutant males (generated using the FRT-FLP-DFS technique). 40% of embryos are able to hatch.

Expression of Apc2^{R1-R5SA.T:Avic\GFP-EGFP} partially suppresses the lethality seen in embryos derived from females carrying homozygous Apc2^g10 Apc^Q8 germline clones and heterozygous double mutant males (generated using the FRT-FLP-DFS technique). 51% of embryos are able to hatch. The remaining progeny exhibit a more severe cuticle phenotype than in the double mutant alone.

Expression of Apc2^{R1-R5SD.T:Avic\GFP-EGFP} partially suppresses the lethality seen in embryos derived from females carrying homozygous Apc2^g10 Apc^Q8 germline clones and heterozygous double mutant males (generated using the FRT-FLP-DFS technique). 49% of embryos are able to hatch. The remaining progeny exhibit a more severe cuticle phenotype than in the double mutant alone.

Expression of Apc2^{R1-R5ExR.T:Avic\GFP-EGFP} fails to suppress the lethality seen in embryos derived from females carrying homozygous Apc2^g10 Apc^Q8 germline clones and heterozygous double mutant males (generated using the FRT-FLP-DFS technique). 21% of embryos are able to hatch. The severity of the cuticle defects in the remaining progeny is similar to the double mutant alone.

Expression of Apc2^{R3-R5ExR.T:Avic\GFP-EGFP} partially suppresses the lethality seen in embryos derived from females carrying homozygous Apc2^g10 Apc^Q8 germline clones and heterozygous double mutant males (generated using the FRT-FLP-DFS technique). 50% of embryos are able to hatch. The remaining progeny exhibit a very weak cuticle phenotype.

Embryos derived from Apc2^g10 Apc^Q8 males crossed to females containing double homozygous germline clones generate naked cuticle. These embryos show approximately 50% lethality (as half the embryos are paternally rescued).

The cuticle defects of embryos derived from Apc2^g10 Apc^Q8 males crossed to females containing double homozygous germline clones are rescued by expression of Apc2^{T:Avic\GFP-EGFP,T:Zzzz\Mito-actA}. The embryonic lethality is partially rescued (19% lethality is seen).

The cuticle defects of embryos derived from Apc2^g10 Apc^Q8 males crossed to females containing double homozygous germline clones are strongly rescued by expression of Apc2^{T:Avic\GFP-EGFP,T:Mmmm\c-Ha-Ras}. The embryonic lethality is partially rescued (41% lethality is seen).

The cuticle defects of embryos derived from Apc2^g10 Apc^Q8 males crossed to females containing double homozygous germline clones are strongly rescued by expression of Apc2^{T:Myr-Src64B,T:Avic\GFP-EGFP}. The embryonic lethality is partially rescued (33% lethality is seen).

The cuticle defects of embryos derived from Apc2^g10 Apc^Q8 males crossed to females containing double homozygous germline clones are partially rescued by expression of Apc2^{T:Avic\GFP-EGFP,T:Hsap\CAAX}. The embryonic lethality is not rescued.

Apc^{WT.T:Avic\GFP}, Apc^{endsatSAMP.T:Avic\GFP} and Apc::Apc2^{APC2+APC1CT.T:Avic\GFP} each strongly rescue the cuticle defects of embryos lacking both maternal and zygotic Apc2[+] function (homozygous Apc2^g10 embryos derived from Apc2^g10 females). The embryonic lethality is also partially rescued (31%, 28% and 21% lethality is seen respectively) and viable adults are recovered.

The cells around the wing pouch in Apc2^g10 Apc^Q8 double mutant third instar larval wing disc clones are apically constricted and invaginated.

Apc2^g10 Apc^Q8 double mutant clone cells generated in the medullar region of third instar larval brains segregate from their neighbours to form cysts. When clones are induced in the medullar neurons the axons do not extend to the medullar neuropil, forming knots in the center of the clones rather than the normal finely fasciculated projections seen in wild type.

Apc^Q8 enhances the embryonic cuticle phenotype seen in Apc2^g10 mutants. In Apc2^g10 Apc^Q8 double mutant embryos all cells are converted to posterior fates. No cells are seen that secrete the denticles normally seen in the anterior cuticle.

Expression of Apc2^{ΔArmRepeats.T:Avic\GFP-EGFP} provides a very weak rescue of the anterior fate loss seen in Apc2^g10 Apc^Q8 maternal/zygotic mutant embryos.

Expression of Apc2^{T:Avic\GFP-EGFP} rescues the anterior fate loss seen in Apc2^g10 Apc^Q8 maternal/zygotic mutant embryos.

Embryos derived from Apc2^g10 Apc^Q8 males crossed to females containing double homozygous germline clones generate naked cuticle. These embryos show approximately 50% lethality (as half the embryos are paternally rescued).

The cuticle defects and lethality of embryos derived from Apc2^g10 Apc^Q8 males crossed to females containing double homozygous germline clones are rescued by expression of Apc2^{WT.T:Avic\GFP-EGFP}, Apc2^{ΔR3.T:Avic\GFP-EGFP} or Apc2^{Δ15.T:Avic\GFP-EGFP}.

The cuticle defects and lethality of embryos derived from Apc2^g10 Apc^Q8 males crossed to females containing double homozygous germline clones are not rescued by Apc2^{KeepR3.T:Avic\GFP-EGFP}, Apc2^{Δ20.T:Avic\GFP-EGFP}, Apc2^{ΔR2.T:Avic\GFP-EGFP}, Apc2^{ΔB.T:Avic\GFP-EGFP} or Apc2^{d40.T:Avic\GFP-EGFP}.

Apc2^{Δ15Δ20.T:Avic\GFP-EGFP} does not rescue the cuticle defects of embryos derived from Apc2^g10 Apc^Q8 males crossed to females containing double homozygous germline clones. Apc2^{Δ15Δ20.T:Avic\GFP-EGFP} may show some dominant negative activity, as 82% embryonic lethality is seen, suggesting that the ability of paternally supplied Apc[+] and Apc2[+] to rescue embryonic lethality is compromised.

Apc2^{ΔSAMP.T:Avic\GFP-EGFP} does not rescue the cuticle defects of embryos derived from Apc2^g10 Apc^Q8 males crossed to females containing double homozygous germline clones. Apc2^{ΔSAMP.T:Avic\GFP-EGFP} may show some dominant negative activity, as 71% embryonic lethality is seen, suggesting that the ability of paternally supplied Apc[+] and Apc2[+] to rescue embryonic lethality is compromised.

23% of embryos derived from females carrying homozygous Apc2^g10 Apc^Q8 germline clones and heterozygous double mutant males (generated using the FRT-FLP-DFS technique) are able to hatch. All of the surviving embryos are maternally double null and zygotically heterozygous for the double-null chromosome, whilst all of the maternally and zygotically mutant embryos died. The embryos display a range of cuticle defects including a reduction in cuticle size due to excess cell death, a hole in the anterior cuticle due to a failure in head involution and the production of excess smooth cuticle at the expense of denticles.

Expression of Apc2^{FL.T:Avic\GFP-EGFP} suppresses the lethality seen in embryos derived from females carrying homozygous Apc2^g10 Apc^Q8 germline clones and heterozygous double mutant males (generated using the FRT-FLP-DFS technique). 46% of embryos are able to hatch. The cuticle defects are also significantly rescued.

Expression of Apc2^{ΔC30.T:Avic\GFP-EGFP} suppresses the lethality seen in embryos derived from females carrying homozygous Apc2^g10 Apc^Q8 germline clones and heterozygous double mutant males (generated using the FRT-FLP-DFS technique). 46% of embryos are able to hatch. The cuticle defects are also significantly rescued.

Expression of Apc2^{N-SAMP.T:Avic\GFP-EGFP} is unable to suppress the lethality seen in embryos derived from females carrying homozygous Apc2^g10 Apc^Q8 germline clones and heterozygous double mutant males (generated using the FRT-FLP-DFS technique). 17% of embryos are able to hatch and the severity of the cuticle defects is similar to in the double mutant alone.

Apc2^g10, Apc^Q8/Apc2^c9 adults shifted to the non-permissive temperature show an increase in BrdU incorporation in the midgut compared to controls maintained at the permissive temperature.

5 days after clone induction in adults, Apc2^g10, Apc^Q8 double mutant clones in the midgut have a significantly increased number of cells per clone compared to controls. There is a significant increase in cell size both in the anterior and posterior midgut, with the average size being greater in the posterior midgut. Intestinal stem cell self-renewal is unaffected at 5 and 10 days after clone induction in the adult.

20 days after clone induction in adults, midguts containing Apc2^g10, Apc^Q8 double mutant clones are associated with gross anatomical changes, including hyperplasia and multilayered cellular masses that distort the luminal surface of the midgut.

Apc2^g10, Apc^Q8 double mutant clones in the adult midgut that are also expressing N^{dsRNA.P.Scer\UAS} under the control of Scer\GAL4^tub often show extensive proliferation and multilayering 10 days after induction in the adult. The number of intestinal stem cells present and the mitotic index of the intestinal stem cells 5 days after induction is significantly higher in Apc2^g10, Apc^Q8 double mutant clones in the adult midgut that are also expressing N^{dsRNA.P.Scer\UAS} under the control of Scer\GAL4^tub compared to clones expressing N^{dsRNA.P.Scer\UAS} under the control of Scer\GAL4^tub in an otherwise wild-type background.

The hyperplasia seen in Apc2^g10 Apc^Q8 double mutant intestinal stem cell clones in the adult midgut is partially suppressed if they are also expressing pan^ΔN.Scer\UAS under the control of Scer\GAL4^tub.

Apical actin in the offspring of Eb1^B13/+; Apc2^g10 mothers appears normal. Furthermore, there is no significant increase in the percentage of incomplete actin rings in these embryos as compared to wild-type.

Embryos in the offspring of dia^k07135/+ ; Apc2^g10 mothers have increased metaphase cap-like actin at the apical surface as compared to single heterozygous controls. The number of incomplete rings is significantly increased in these embryos.

The photoreceptor cell apoptosis phenotype seen in Apc^Q8 homozygotes is not suppressed by Apc2^g10/+.

There is no difference in the frequency of mispositioned oocytes between wild type and Apc2^g10, Apc^Q8 double mutant mutant germlines.

Apc2^g10, Apc^Q8 maternal/zygotic double mutant embryos show no defects in epithelial structure and do not show a total disruption of the cuticle.

Apc2^g10, Apc^Q8 maternally mutant syncytial embryos show similar levels (35 vs 37%) of cortical nuclei movement to the anterior to Apc2^g10 syncytial embryos.

Apc2^g10, Apc^Q8 maternally mutant syncytial embryos do not have significant defects in overall spindle morphology or orientation but do show a slight but significant lengthening of the pole-to-pole distance. These double mutants show normal symmetric cell division.

Apc2^g10, Apc^Q8 double homozygotes die as second instar larvae.

Apc2^g10 Apc^Q8 double zygotic mutants are embryonic viable and exhibit a wild-type cuticle pattern, but die as larvae.