(a) Immunostaining of brains from 10-day-old elavGS>UAS-BNIP3 flies with or without RU486-mediated transgene induction from day 5 onward, showing BNIP3 expression level (green channel, anti-HA) and nuclear DNA (blue channel, stained with DAPI). Scale bar is 50 μm.
(b) Quantification of BNIP3 expression level in brain as shown in (a). n = 10 biologically independent animals per condition. ***p<0.0001; unpaired t test. Data are presented as scatter plots overlaying mean values +/− SEM.
(c) Western blot detection of BNIP3 transgene induction in the heads of day 14 elavGS>UAS-BNIP3 flies with or without 9 days of RU486 treatment. n = 5 biological replicates per condition with 10 flies pooled per replicate.
(d) Immunostaining of brains from young (10-day-old) elavGS>UAS-BNIP3 flies with or without RU486-mediated transgene induction from day 5 onward, showing mitochondrial morphology (green channel, anti-ATP5a) and BNIP3 (red channel). Arrows and outlines indicate sites of colocalization. Scale bar is 3 μm. RU486 was provided in the media at a concentration of 5 μg/ml. Images are representative of 5 samples treated with RU and 6 samples provided vehicle.