Hsp83 gene expression level in larvae of three different D. melanogaster populations. Fly lines homozygous for the Hsp83 mutant (Hsp83P/P, red) and wild-type (Hsp83+/+, blue) strains were reared at two different temperatures (25°C and 28°C) for one generation before RNA was extracted. Flies with each genotype were isolated from three populations from Ivory Coast, Okayama, and Tokyo, respectively, as indicated. Note that Hsp83 expression in flies maintained at 28°C is downregulated compared with flies maintained at 25°C, which stands in contrast with Hsp83 expression in heat-shocked flies . Hsp83 mRNA was purified from at least 20 early third-instar larvae, and quantified using real-time PCR. The expression of the housekeeping gene actin88F was used as an endogenous loading control. Expression levels of Hsp83 are measured relative to the expression of actin88F. Triplicate PCRs were performed for each mRNA sample.
Chen and Wagner BMC Evolutionary Biology 2012 12:25 doi:10.1186/1471-2148-12-25