Extended Data Fig. 2: Screening of the female-specific CYP4PC1 by RNA-seq and its expression profiling.
From: A single gene integrates sex and hormone regulators into sexual attractiveness
a, Distribution of cuticular C29 methyl ketone titre in sexually mature females. The antennae and abdominal integument (in red text and bar) were used for RNA-seq analyses. n = 3 biological replicates. Different letters indicate statistically significant differences between groups using Welch’s ANOVA (Games-Howell multiple comparisons test, P < 0.05). Data are mean ± s.e.m. b, The numbers of total CYP genes identified in the antennae and abdominal integument, and those upregulated in sexually mature females compared to males. The FPKM values of the up-regulated CYPs are available in Supplementary Table 1, 2. c, Genomic organization of CYP4PC1 and characteristic P450 motifs identified from the deduced protein sequence. The numbers nearby indicate the length of each exon or intron (base pair, bp), and conserved motifs typical of P450 are marked with colour-filled boxes. d, Developmental patterns of CYP4PC1 expression in various tissues across sexual maturation. n = 4 biological replicates. Data are mean ± s.e.m. e, Gel electrophoresis verification of a 1,619 bp fragment containing the complete coding sequence of CYP4PC1, as amplified from various tissues by RT-PCR. A 104 bp fragment of actin-5c was used as a loading control. M, DNA marker. -, negative control with no DNA template. f, Western blotting validation of the CYP4PC1 antibody using antennal protein extracts and RNAi experiment. The red frame indicates immune signals of the endogenous CYP4PC1 protein, with a predicted molecular weight of 58 kDa. M, protein marker. The lane of protein markers was spliced from the white light image of the same membrane. g, Dose-dependent CYP4PC1 protein level in various tissues resulted from a gradient of total protein loaded, as determined by western blotting.
