Figure 2.

Generation of rdhB¹ and effects of mutation on chromophore and Rh1 production during the pupal period. A, Schematic illustration of the rdhB (CG7077) gene and generation of the rdhB knock-out (rdhB¹) by ends-out homologous recombination. B, Western blot containing head extracts from the indicated flies probed with anti-RDHB and reprobed with anti-Tubulin (Tub). C, Western blot of head extracts from ≤1-d-old wild-type and rdhB¹. The flies were raised from the second instar larval stage on retinoid-free medium supplemented with 5 mm β-carotene or 500 μm all-trans-retinal or all-trans-retinol. The flies were kept under a 12 h light/dark cycle. D, Quantification of Western blot results shown in C. n ≥ 3. E, Decreased chromophore levels in rdhB¹ flies fed all-trans-retinal or all-trans-retinol. The flies were ≤1 d old and raised as indicated in C. n = 3. The concentration of 3-OH-11-cis-retinal and 3-OH-all-trans-retinal were measured by HPLC. Error bars represent ±SEMs. *p < 0.05. Unpaired Student's t tests.