Figure 8.

mTORC2 Regulates PDGFR Expression via FOXO3a in Smooth Muscle Cells

Pharmacological inhibition of mTORC2 up-regulates the nuclear protein expression of Forkhead box O3a (FOXO3a) in human PASMCs. (A) Representative images showing Western blot analyses on FOXO3a in the total, nuclear, and cytoplasmic proteins idolated from human PASMCs treated (for 24 h) with vehicle (Cont), Rap (50 nM), and KU 0063794 (KU, 200 nM) (a). Summarized data (mean ± SE) showing the total (Kruskal-Wallis test, p = 0.01; n = 4 in each group), nuclear (Kruskal-Wallis test, p = 0.01; n = 5 in each group), and cytoplasmic (Kruskal-Wallis test, p = 0.06; n = 5 in each group) protein levels of FOXO3a in PASMCs treated with vehicle (Control; blue bars), Rap (green bars), and KU (red bars) (b). Dunn test, *p < 0.05, **p < 0.01 versus Control; #p < 0.05 versus rapamycin. (B) Representative immunofluorescence images showing cell nuclei (DAPI; blue) and FOXO3a (dark green) in human PASMCs treated (for 24 h) with vehicle (Control), Rap (50 nM), and KU (200 nM) (a). The fluorescence intensity of FOXO3a along the line across a PASMC (b) treated with vehicle (Control), Rap, and KU. Summarized data (mean ± SE) showing the mean fluorescence intensity of the nuclear FOXO3a (c) in Control (n = 24), Rap-treated (n = 18), and KU-treated (n = 21) PASMCs; the value was calculated as the ratio of the nuclear intensity of FOXO3a to the cytoplasmic fluorescence intensity of FOXO3a. Kruskal-Wallis test, p < 0.001 and Dunn test, ***p < 0.001 versus Control; ^###p < 0.001 versus rapamycin. Abbreviations as in Figures 1 and 7.