Bao and Xu BMC Genomics 2011, 12:224 http://www.biomedcentral.com/1471-2164/12/224 Page 7 of 14 Figure 4 Developmental expression patterns of genes isolated by SSH. Expression pattern of four genes during the early stage of Helicoverpa armigera pupae. Total RNA and protein were extracted from brains and SGs or brain–SG complexes of diapause- and nondiapause- destined pupae. (A) Expression of genes measured by RT-PCR. The cDNAs of the four genes were amplified by PCR with 20–24 cycles; actin was amplified with 20 cycles. (B) Western blot analysis. Total protein (20 μg) was separated and incubated with polyclonal antibodies (Har-SUMO, 1:1000; Har-MnSOD, 1:3000; Har-Sericotropin, 1:2000; Har-TCTP, 1:3000). The numbers (0–5) represent the days after pupation, and actin was used as an internal standard. NP, nondiapause-destined pupae; DP, diapause-destined pupae.