Source was embryos of wild-type fly line; protein produced from endogenous gene.

The lola zf5 (lola-PD) 105 kDa isoform was immunoprecipitated using the isoform-specific 7F1 monoclonal antibody. Other coimmunoprecipitated isoforms of lola (of different molecular weight) were detected using a polyclonal lola antiserum.

From an analysis of many TF-ChIP data sets, TF-TF interactions were predicted on the basis of regular DNA sequence motif arrangements. 27 predicted TF-TF interactions were tested using a pull down assay that measures the \'Luminescence Intensity Ratio\'. A cut-off of 7 was used, well above values for six negative controls.

Homotypic interactions confirmed for lola-PA, lola-PD and lola-PQ isoforms.

Interaction in vitro; bait produced by coupled in vitro translation; prey produced by coupled in vitro translation.

Two-hybrid system: yeast GAL4-BD/GAL4-AD

Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein (prey was previously cloned reagent).

Untagged lola forms a 5.7 mer with an apparent molecular mass of 86.3 kDa and MBP tagged lola forms a 5.6 mer with an apparent MW of 315 kDa, as determined by multi-angle light scattering.

Interaction in vitro; protein produced as a recombinant fusion protein in bacterial system.

MW values were determined from column calibration with standard proteins in addition to staining. lola forms multimers. Mutations in hydrophobic residues to break H-bonds (proline), increase polarity (lysine) or traditional alanine, all decreased hexamer formation.

Interaction in vitro; protein produced as a recombinant fusion protein in bacterial system.

Interaction in vitro; protein produced as a recombinant fusion protein in bacterial system.

SAXS data support that the prevalent oligomer state in the solution of lola TTK-type BTB domain is the hexamer, in accordance with the CryoEM data.

Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein (prey was previously cloned reagent).

Two-hybrid system: yeast GAL4-BD/GAL4-AD