Rhabdomeral localization of Rh1 and TRP is not affected in the <i>ttd14</i>^<i>P75L</i> mutant.

<p>(A) Water immersion microscopy images of the Rh1-eGFP and TRP-eGFP fluorescence in the eyes of wild type flies (wild type) and <i>ttd14</i>^<i>P75L</i> mutant eye clones (<i>ttd14</i>^<i>P75L</i>). Flies were kept in constant darkness for 1 day (upper row), 7 days (middle row) or 28 days (lower row). Scale bar: 10 μm. (B) Analysis of Rh1-eGFP and TRP localization on cross sections through ommatidia of wild type flies (wild type) and <i>ttd14</i>^<i>P75L</i> mutant eye clones (<i>ttd14</i>^<i>P75L</i>), kept in darkness for 7 d. Rh1-eGFP localization was detected by its eGFP fluorescence (green, upper row), TRP localization was detected by an anti-TRP-antibody (green, upper row). The actin cytoskeleton of rhabdomeres was labeled with Alexa Fluor 546-coupled phalloidin (red, middle row). Overlay of red and green fluorescence appears yellow in the merged panels. Scale bar: 5 μm. (C) Immunoblot assessing Rh1 and TRP from heads of wild type flies (wild type, lanes 1–3), <i>ttd14</i>^<i>P75L</i> mutant flies (<i>ttd14</i>^<i>P75L</i>, lanes 4–6) and <i>ttd14</i>^<i>P75L</i><i>; Rh1</i>><i>ttd14-myc</i> flies (rescue, lane 7–9). Protein from 0.5 heads was loaded per lane. Flies were kept in darkness (lanes 1,4,7), exposed to white light (1800 lux) for 20 hours (lanes 2,5,8), or exposed to white light for 20 hours followed by a 6 hour recovery in the dark (lanes 3,6,9). The size of molecular weight markers in kilo Dalton is indicated at the left. (D) Quantification of the Rh1 levels normalized to TRP. Error bars show SEM (n = 4). Rh1 levels decrease after illumination due to lysosomal degradation of internalized Rh1. No significant differences were found between wild type, <i>ttd14</i>^<i>P75L</i> mutant and rescue flies at any light condition.