Figure 5: Expression profiles of the members of the CPR gene families (RR1, RR2 and RR3) as determined by RT-PCR and RT-qPCR.

(A) Expression of RR-1 gene-annotated endocuticle protein genes in different tissues on day 6 of 5^th instar nymphs, as detected by RT-PCR (28 cycles). Different tissues: Integument, head, leg, pronotum, goad, gut, Malpighian tubules, fat body, wing pad; (B) Expression of RR-1 genes in the pronotum after molting from 4^th instar nymphs to 5^th instar nymphs at 0 h, 36 h, 72 h and 96 h, as detected by RT-qPCR. Heat map showing the relative expression level during different stages of RR-1 genes. The colors in the map display the relative values of all tiles within the 4 given developmental stages. Blue indicates the lowest expression, green indicates intermediate expression, and red indicates the highest expression. The color scale bar is shown on the right of the figure. (C) Expression of RR-2 gene-annotated adult cuticle protein genes in different tissues on day 2 of adults as detected by RT-PCR (28 cycles). Different tissues: Integument, leg, muscle, tentacle, goad, gut, Malpighian tubules, fat body, wing. (D) Expression of RR-2 genes in whole body, wing pad or wing from embryo, 1, 2^nd, 3^rd, 4^th, 5^th instar nymphs to the adults as detected by RT-qPCR. (E) Expression of RR-2 genes in the wing of 5^th instar nymphs at different days as detected by RT-qPCR. N4D1, N4D3, N4D5: Day 1, Day 3, Day 5 of 5^th instar nymphs. (F) Expression of RR-3 genes in different tissues on day 6 of 5^th instar nymphs as detected by RT-PCR (28 cycles). Different tissues: Integument, head, leg, goad, gut, Malpighian tubules, fat body, wing pad, pronotum; (G) Expression of RR-3 genes in the pronotum after molting from 4^th instar nymphs to 5^th instar nymphs at 0 h, 36 h, 72 h and 96 h, as detected by RT-qPCR. β-actin was used as the reference control for RT-PCR (24 cycles) and RT-qPCR, respectively. All data are reported as means ± SE of three independent biological replications. The electrophoresis image were obtained by using the gel imaging analysis system (Bio-Rad, USA). The full-length gels are presented in Supplementary Figs 1–4, Figs 2–4 and Figs 3–4, respectively.