Recessive toxicity of FicE247G expression.
A, structure of the Drosophila fic gene and protein with the regulatory Glu-247, dimerization Ile-271, and catalytic His-375 residues. The CRISPR/Cas9-induced fic30C mutation induces a 101-base pair deletion, generating an early stop codon N-terminal to the conserved TPR domain. B, a representative Western blot shows expression of UAS-Fic transgenes (wild type, H375A, and E247G) under control of the ubiquitous Da-Gal4 driver in a wild-type background. Quantification of Western blots from three separate experiments showed no significant difference in expression of the UAS-Fic transgenes. C, Da-Gal4 homozygous females were crossed to the indicated UAS-Fic/TM6b, Hu males either in a wild-type or fic30C background. The graph represents percentage of progeny inheriting the indicated UAS-Fic transgene together with the Gal4 driver. The expected Mendelian percentage is 50%. Bars, mean of three independent crosses; dots, values for independent crosses; error bars, S.D. The total number of flies scored for each genotype was at least 150. D, representative SEM images of eyes expressing the indicated UAS-Fic transgene with LongGMR-Gal4 in a wild-type or fic30C background. Scale bar, 100 μm. E, representative SEM images of eyes expressing the indicated UAS-Fic transgene along with LongGMR-Gal4, UAS-FicE247G in a fic30C background. Scale bar, 100 μm.