Fig. 10.

PKA and PKC activity in vascular smooth muscle cells (VSMCs) isolated from wild-type (WT) and gravin-t/t mice in response to PDGF stimulation. Western blot analysis of homogenates isolated from aortic VSMCs from WT and gravin-t/t mice using antibodies to the phosphorylated (Thr197) PKA catalytic subunit (p-PKA; A), phosphorylated (Ser/Thr) PKA-dependent substrate (p-PKA substrates; B), phosphorylated (Thr638/641) PKCα/βII (p-PKC; C), phosphorylated (Ser) PKC-dependent substrate (p-PKC substrates; D), phosphorylated (Thr202/Tyr204) Erk1/2 (p-Erk1/2; E), and phosphorylated (Ser⁴⁷³) Akt (p-Akt; F), in response to 10 ng/mL PDGF stimulation as well as in the absence or presence of membrane-permeable selective calcium chelator, BAPTA-AM. In each of the images shown in A–F, the top shows a representative Western bot and the bottom shows the corresponding Western blot quantification of the phospho-protein expression. Results are presented as the mean ± SE. n = 5 (WT, Gravin-t/t), n = 4 (WT PDGF, Gravin-t/t PDGF, WT PDGF BAPTA-AM, Gravin-t/t PDGF BAPTA-AM) (A); n = 5 (WT, Gravin-t/t), n = 7 (WT PDGF), n = 6 (Gravin-t/t PDGF), n = 5 (WT PDGF BAPTA-AM, Gravin-t/t PDGF BAPTA-AM) (B); n = 4 (WT, Gravin-t/t, WT PDGF, Gravin-t/t PDGF), n = 3 (WT PDGF BAPTA-AM, Gravin-t/t PDGF BAPTA-AM) (C); n = 5 (WT), n = 4 (Gravin-t/t), n = 4 (WT PDGF), n = 6 (Gravin-t/t PDGF), n = 5 (WT PDGF BAPTA-AM), n = 4 (Gravin-t/t PDGF BAPTA-AM) (D); n = 4 (WT), n = 3 (Gravin-t/t), n = 5 (WT PDGF), n = 4 (Gravin-t/t PDGF, WT PDGF BAPTA-AM, Gravin-t/t PDGF BAPTA-AM) (E); n = 4 (WT, Gravin-t/t) n = 5 WT PDGF, Gravin-t/t PDGF), n = 3 (WT PDGF BAPTA-AM, Gravin-t/t PDGF BAPTA-AM) (F). *P < 0.05, **P < 0.01, and ***P < 0.0001. Comparisons between two groups were determined by an unpaired two-tailed Student’s t test, and comparisons between multiple groups were determined by one-way ANOVA followed by post hoc Tukey test. MM, molecular mass.