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. Author manuscript; available in PMC: 2018 Nov 10.
Published in final edited form as: Circ Res. 2017 Oct 11;121(11):1251–1262. doi: 10.1161/CIRCRESAHA.117.311819

Figure 4. Lactate-induced phenotypic modulation of hiPSC-vSMCs is mediated by NDRG3.

Figure 4

(A–B) hiPSC-derived vSMCs were cultured in L+ medium, in L− medium, or in L− medium under hypoxic conditions (L−vSMCh); then, (A) the levels of NDRG3 protein (left) and mRNA (right) were evaluated via Western blot and quantitative RT-PCR (n=3), respectively, and (B) protein levels of Raf, phosphorylated Raf (P-Raf), (ERK), and phosphorylated ERK (P-ERK) were evaluated via Western blot. (C-F) vSMCs were transfected with NDRG3 iRNA or scrambled (control) iRNA. (C) NDRG3 protein levels in NDRG3-iRNA–transfected and control-iRNA–transfected vSMCs were evaluated via Western blot. (D-E) NDRG3-iRNA–transfected and control-iRNA–transfected SMCs were cultured in L+ medium; then, (D) proliferation was evaluated via proliferation assay (n=3) and (E) collagen 1 production was measured via ELISA (n=3). (F) NDRG3-iRNA–transfected and control-iRNA–transfected vSMCs were cultured in L+ medium for 3 days; mRNA levels of MYH11, collagen 1 (Col 1), and vimentin (VMT) were measured via quantitative RT-PCR before and after the culture period (n=3). **P<0.01, ***P<0.001 for all panels.