Third instar larval eye-antennal imaginal disc clones co-expressing dlg1GD4689 and Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.CoinFLP-FRT, in combination with Dicer-2 for efficient RNAi, form invasive tumors and induce autophagosomes in a cell non-autonomous manner in neighboring disc tissue (assessed by a Atg8a fluorescence reporter), as compared to controls; both the tumor formation and the cell non-autonomous autophagosome induction are suppressed by the additional clonal co-expression of either sdJF02514 or ykiHMS00041; the volume of these tumors is also significantly reduced by the co-expression of either slifUY681 or Pi3K92ED954A.Scer\UAS.T:Hsap\MYC under the control of Scer\GAL4Act5C.CoinFLP-FRT; the cell non-autonomous increase in autophagosomes is also suppressed by the clonal co-expression of bskDN.Scer\UAS.cUa, but not of ImpL2dsRNA.Scer\UAS.cUa or Pi3K92ED954A.Scer\UAS.T:Hsap\MYC.
The co-expression of Ras85DV12.UAS and dlg1GD4689 under the control of Scer\GAL4ap-md544 induces wing disc neoplasia which, when transplanted, can form solid tumors in the host; there is amplification of the resident myoblast population, as well as accumulation of circulating hemocytes on the wing disc. Myoblast ablation (by expressing rprlexAop.cSa under the control of Ecol\lexAGMR15B03) does not suppress the tumor-like growth.
The co-expression of Ras85DV12.UAS and dlg1GD4689 under the control of Scer\GAL4hh.PU induces wing disc tumor-like growth, with increased mitotic index.; myoblast ablation (by expressing rprlexAop.cSa under the control of Ecol\lexAGMR15B03) does not suppress the tumor-like growth. Similar tumor induction occurs if expression is induced in wing disc transplants (expression induction is controlled by Gal80[ts] and temperature-shift); hemocyte ablation on the host (by expressing rprUAS.C under the control of Scer\GAL4He.PZ) together with myoblast ablation on the implant (by expressing rprUAS.C under the control of Ecol\lexAGMR15B03) does not suppress the tumor-like growth.
The tumor-like overgrowth of MARCM clones in larval antennal disc expressing dlg1GD4689 under the control of Scer\GAL4Dll-md23 is not significantly worsened by cnoR2 homozygosity but the cellular and tissue morphology and disorganization are exacerbated.
Expression of ImpL2GD6004 within the tumors of Scer\GAL4ey.PU>Ras85DV12.Scer\UAS,dlg1GD4689 origin significantly ameliorates the peripheral tissue wasting phenotypes. Hosts bearing dlg1GD4689, Ras85DV12.Scer\UAS, ImpL2GD6004 tumors show increased abdominal fat body mass. Muscle function assays further reveal improvements in both climbing ability and speed. There is a significant rescue of ovariole health, leading to restoration of egg production.
Co-expression of Pi3K92EGD11228 suppresses tumour growth in dlg1GD4689 MARCM third instar imaginal disc clones, although this suppression is weaker than that seen upon expression of Pi3K92EGD11228 in Ras85DV12.Scer\UAS-dlg1GD4689 MARCM third instar imaginal disc clones.
Knockdown of either Pi3K92E or Akt1 in Ras85DV12.Scer\UAS-dlg1GD4689 cells, through expression of either Pi3K92EGD11228 or Akt1GD1361, causes a complete suppression of neoplastic growth and even causes a near-complete absence of mutant eye cells. Only small structures, which contain few cells, remain.
MARCM Ras85DV12.Scer\UAS-dlg1GD4689 expressing clones are dramatically smaller in a Pi3K92E1C1 homozygous mutant than in a wild-type background, indicating that a loss of Pi3K92E signaling interferes with Ras85DV12.Scer\UAS-dlg1GD4689 tumour growth.
Expression of Ras85DV12.Scer\UAS and dlg1GD4689 in the posterior compartment of wing discs, under the control of Scer\GAL4en.PU results in an enlarged posterior wing disc compartment. These wing discs exhibit high levels of cell proliferation.
Co-expression of Pi3K92EGD11228 together with Ras85DV12.Scer\UAS and dlg1GD4689, under the control of Scer\GAL4en.PU, strongly reduces the size of the Ras85DV12.Scer\UAS-dlg1GD4689 tumours in the posterior wing disc compartment. There is an almost complete absence of cellular proliferation in this compartment.
Wing disc compartments expressing Ras85DV12.Scer\UAS, dlg1GD4689, Pi3K92EGD11228, together with CycEScer\UAS.cRa and stghs.PE2 show a rescue in compartment size and high levels of His3-positive cells.
The moderate rough eye phenotype and reduced adult eye size characteristic for flies expressing Scer\GAL4GMR.PU under the control of Hsap\MAPTV337M.Scer\UAS is significantly worsened by co-expression of dlg1GD4689 (together with UAS-Dicer2 to enhance RNAi efficiency), exacerbating the rough appearance.