Figure 3.

SGK3 enhanced high phosphate-induced expression of Pit-1 in cultured VSMCs. A-D. Mouse VSMCs transiently transfected with SGK3-S486D plasmid or SGK3 siRNA. The transcription levels of Pit-1 and SGK3 measured by qPCR (A, C). The protein levels of Pit-1 and SGK3 were measured by western blot (B, D). E. Mouse VSMCs treated with SGK3-PROTAC1 for 24 h. Immunoblot of Pit-1 and SGK3 cleavage of cell lysates. F. Mouse VSMCs were transfected with scramble or SGK3 shRNA for 72 h. The protein levels of Pit-1 and SGK3 were measured by western blot. *P < 0.05, **P < 0.01, ***P < 0.001 vs. control group. G-J. Mouse VSMCs transiently transfected with SGK3 siRNA or SGK3-S486D plasmid, in presence or absence of 3 mM Pi. The protein levels of Pit-1 and SGK3 were measured by western blot (G, I). Pi uptake levels were determined from cell lysates by Pi Assay Kit (H, J). *P < 0.05, **P < 0.01, ***P < 0.001 vs. control group; ^#P < 0.05, ^##P < 0.01, ^###P < 0.001 vs. Pi group.