Fig. 2: PG-specific depletion of Evi5 causes iron-deficiency.

A Red autofluorescence in ring glands from PG > w¹¹¹⁸ (control) and PG >FLP;Evi5^FRT larvae reared on a normal diet (no suppl. = no supplement), or on media supplemented with FAC (iron), Zinc-Protoporphyrin IX (ZnPP), Hemin and 20-Hydroxyecdysone (20E). The dotted lines show boundaries of the brain and the RG under UV and brightfield (BF) light. Scale bar = 250 μm. B Expression of ALAS in RGs isolated from PG >FLP;Evi5^FRT larvae relative to the control (shown as dotted line = 1) after being raised on different foods. Fold changes 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 the error bars denotes the average. Asterisks are P-values obtained via a two-sided, paired Student’s t-test (***p < 0.001). C Survival rates of PG>Ppox^IR on normal diet (no suppl.), Hemin- and ZnPP-supplemented media. D Pupariation timing of PG >FLP;Evi5^FRT animals when reared on different media. E Pupal lethality of PG >FLP;Evi5^FRT animals when reared on different media. L3: 3rd instar larvae, P: pupae, Ad: adults. Significance letters are based on P-values obtained via a two-sided, paired Student’s t-test (***p < 0.001). In C-E, error bars represent standard error, with the center denoting the average. In (C–E), three biological replicates were used, with each sample consisting of 50 individuals. F X-Ray Fluorescence Microscopy (XRF) images of BRGCs from control and PG >FLP;Evi5^FRT animals. At the 60 and 74 h, the control animals underwent puparium formation and are no longer comparable to the PG >FLP;Evi5^FRT larvae. G Quantification of iron distribution in ring glands (red dotted line) vs. CNS (black dotted line) from control and PG >FLP;Evi5^FRT animals. Image scale bars represent 80–100 µm. The color scale bar is log2-based and represents tissue iron levels. Significance in E and G is based on ANOVA and letter differences indicate a two-sided P-value of p < 0.05. Error bars denote standard deviation of number pixels in selected areas, with the center denoting the average. Source data are provided in the source data file.