Fig. 3: PG-specific RNAi screen of Rab, Rab-GAP, Rab5-GEF and Rab11-GEF genes.

A Time required for control (PG >w¹¹¹⁸) and PG>Rab5^IR animals to reach pupariation. Error bars represent standard error of three biological replicates (each = 50), with the center indicating the average. B Survival analysis of PG>Rab11^IR animals. Error bars are standard error of the mean for three biological replicates (each = 50) and asterisks represent the P-value obtained by the two-sided Student’s t-test (***p < 0.001). L3: 3^rd instar larvae, P: pupae, Ad: adults. IR = RNAi line. C Red autofluorescence of ring glands obtained from PG>Rab5^IR and PG>Rab11^IR animals. UV: ultraviolet light, BF: bright field. Scale bar = 250 μm. D qPCR analysis of the ALAS in ring glands of PG>Rab5^IR and PG>Rab11^IR animals. E Relative expression of Rab5 and Rab11 in ring glands isolated from PG >FLP;Evi5^FRT animals based on qPCR analysis. In (3D, 3E), fold changes were calculated with ddCT method and error bars denote 95% confidence intervals, both are based on three biological replicates (each tested in triplicate). The center of error bars denotes the average. Asterisks are P-values obtained via a two-sided, paired Student’s t-test (***p < 0.001). F PG-specific expression of Evi5^WT-mVenus and Evi5^R160A-mVenus cDNAs (green label) in either wild type or Evi5 mutant (PG > FLP;Evi5^FRT) backgrounds. The dotted lines outline the ring gland and attached brain section under brightfield (BF), green (mVenus) and Red (UV) channels. Scale bar = 150 μm. G. H Heatmaps showing pupariation profiles for lines expressing PG-specific RNAi targeting Drosophila Rab, Rab-GAP, Rab5-GEF and Rab11-GEF genes. Shown is the percentage of animals that have reached puparium formation at a given time point. Gene names in red (Evi5, Rab5 and Rab11) indicate the presence of red autofluorescing ring glands in the corresponding RNAi line. Rab11^CA is a constitutively active form of Rab11. Green gene names represent Rab5-GEFs (nucleotide exchange factors), whereas blue names represent Rab11-GEF genes. Source data are provided in the source data file.