Figure 2.

Overexpression of KLF4 inhibited the activation of mTOR. VSMCs were co-infected with AdKLF4 and AdtTA (20 MOI) and maintained in the medium with or without tetracycline (Tc; 0.1 µg·mL⁻¹). (A) Nuclear protein lysates were immunoblotted with antibodies against KLF4 or histone H3 as an internal control. (B) Immunofluorescent staining was performed using a primary antibody against KLF4 and detected with a rhodamine-conjugated secondary antibody. Nuclei were counterstained with Hoechst 33258. (C) VSMCs were treated with PDGF-BB (20 ng·mL⁻¹) for the indicated time, and total proteins were immunoblotted with antibodies against mTOR, p70S6K, 4EBP1 or their phosphorylated forms. Data shown are representative of three independent experiments. (D) Nuclear protein lysates were immunoblotted with antibody against p27^kip1 and histone H3. Data are representative of three independent experiments. **P < 0.01, *P < 0.05, significantly different from VSMCs infected with AdtTA and AdKLF4 in the presence of Tc.