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. 2018 May 23;9(6):612. doi: 10.1038/s41419-018-0598-6

Fig. 6. Knockdown of Sec61α inhibits Nogo-C- and TGF-β1-mediated fibrotic responses in cardiac fibroblasts.

Fig. 6

Fig. 6

a Western blot and average date showing Sec61α and Nogo-C protein levels in cardiac fibroblasts transfected with Ad-sh-Sec61α or Ad-scramble for 72 h. n = 3 independent experiments. b Western blot and average date showing Nogo-C and Sec61α protein levels in cardiac fibroblasts transfected with Ad-sh-Nogo-C or Ad-scramble for 72 h. n = 3 independent experiments. c Immunofluorescence staining showing the co-localization of Nogo-C with Sec61α in cardiac fibroblasts with/without TGF-β1 or AngII stimulation. Scale bar = 10 μm. d CoIP of Nogo-C protein with Sec61α protein in cardiac fibroblasts. e Ubiquitination level of Sec61α protein in cardiac fibroblasts transfected with Ad-Nogo-C or Ad-LacZ in the presence of MG132 (10 μM). f Cytosolic Ca2+ concentration in cardiac fibroblasts transfected with Ad-Nogo-C with/without co-transfection of Ad-sh-Sec61α. g Western blot and average date showing levels of profibrogenic proteins TGF-β1, CTGF, FN, and collagen type I in cardiac fibroblasts transfected with Ad-Nogo-C with/without co-transfection of Ad-sh-Sec61α. h Profibrogenic proteins and Nogo-C in cardiac fibroblasts treated with TGF-β1 (10 ng/ml) with/without knockdown of Sec61α by Ad-sh-Sec61α transfection. n = 3 independent experiments. i, j Immunofluorescence staining of FN (i) or α-SMA (j) in cardiac fibroblasts in response to TGF-β1 (10 ng/ml) stimulation in the presence/absence of Ad-sh-Sec61α infection. Scale bar = 25 μm. *P < 0.05, **P < 0.01, ***P < 0.001 vs Ad-LacZ or control group. #P < 0.05, ##P < 0.01, ###P < 0.001 vs Ad-Nogo-C or TGF-β1 group