Fig. 3. Relative quantification of the effect of 20E on CqVg expression in vivo and in primary hepatocyte cell culture. (A) Intact and XO–SG-extirpated crayfish were injected daily with either 20E or carrier until they reached ecdysis. They were then sacrificed, and RNA was extracted for evaluation of CqVg expression levels. The control intact crayfish was not subjected to any injection. Different letters indicate statistical significance (P<0.05). (B) A 24 h primary hepatocyte cell culture from an intermolt male was subjected to four different concentrations of 20E: 1, 10, 100 and 1000 pg µl^–1. In the carrier sample no 20E was added. The positive control was a 24 h female primary hepatocytes cell culture with no addition of 20E (Female). Different letters indicate statistical significance (P<0.05).