Figure 5.
Target gene knockdown reduces maximal adenylyl cyclase activity in CAD VN-AC5/D2L cells. siRNA knockdown of the target genes reduced the maximal forskolin-stimulated AC activity in CAD VN-AC5/D2L cells (A). The ability of the Gαi/o-coupled D2 receptor to acutely inhibit AC activity remained intact following siRNA knockdown (B). siRNA knockdown of the target genes resulted in a reduction of the maximal sensitization response, with no difference in the potency of quinpirole to promote the development of sensitization (C). Experiments (n = 3) were performed in duplicate with SEM shown. Biochemical validation of siRNA knockdown was carried out by western blotting (D,E). Cell fractions from mock or siRNA-treated cells were subjected to western blot analysis using target antibodies for (D) PPP2CB and (E) NAPA, as well as listed loading controls. Data were standardized to mock transfection (left) and representative immunoblots (right). Statistical analysis was performed (t-test), *** p < 0.001.