Additional file 6.

Figure S3. Experimental design for continuous inbreeding of flies at 25°C and 28°C. Flies from 7 isofemale lines for each genotype (wildtype Hsp83+/+ vs. mutant Hsp83P/P) were subject to the same inbreeding procedure in parallel. In each generation, four 3-day-old virgin females and four 3-day-old males of an isofemale line were mated in a vial to generate the next generation. The 8 flies in the vial were transferred to a new vial with fresh food every other day for a total of 30 days. Adults eclosing from each vial were censused every other day until eclosion ceased. At 25°C inbreeding continued for 4 generations. At 28°C inbreeding continued for 3 generations for wild-type flies, and for 2 generations of mutant flies, because the flies ceased to lay eggs in the second generation. Fly lines isogenized from each population were subject to the same inbreeding procedure.

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Chen and Wagner BMC Evolutionary Biology 2012 12:25   doi:10.1186/1471-2148-12-25