Fig 6. roh mutation results in Rh1 decrease.

(A) GFP fluorescence of one-day old wild-type (left panel) and roh ^EY04039 (right panel) flies showing a reduction in Rh1-GFP. Scale bar, 50 μm. (B) Western blotting showing decreased Rh1 levels in the roh ^EY04039 mutant (ey-flp Rh1::GFP; FRT42D roh ^EY04039 / FRT42D GMR-hid CL); the INAD and TRP levels are not affected compared to the wild-type. The reduction of Rh1 levels could be rescued in roh ^ex (ey-flp Rh1::GFP; FRT42D roh ^ex /FRT42D GMR-hid CL) and roh ^EY04039;GMR>roh (ey-flp Rh1::GFP; FRT42D roh ^EY04039 / FRT42D GMR-hid CL;GMR-gal4/UAS-roh) flies. Flies less than 1 day old were used. (C) Quantification of relative Rh1 level in various genotypes: wt, ninaA ¹, roh ^EY04039, roh ^ex and roh ^EY04039;GMR>roh. The Rh1 levels were normalized to tubulin. (D) Quantitative real-time PCR of wild-type (ey-flp Rh1::GFP; FRT42D/ FRT42D GMR-hid CL) and roh ^EY04039 fly head with rh1-specific primers. The results are normalized to the expression level of gpdh. The graphs represent the means ± SD of three independent experiments. Error bars represent the SD. (E) Average rhabdomere numbers per ommatidia of the roh ^EY04039 mutant under the indicated conditions. Each data point was based on examination of >60 ommatidia from >3 flies. Error bars represent the SDs. Asterisks indicate statistically-significant differences (one-way ANOVA and post-hoc Dunnett’s test; ns: not significant, **p < 0.01).