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. 2010 Mar;176(3):1139–1147. doi: 10.2353/ajpath.2010.090459

Figure 7.

Figure 7

Effect of proteasome inhibitor MG132 on SnoN expression, Smad3 signaling, and TGF-β-dependent PAI-1 expression. A and B: HK-2 cells were incubated with control medium or 1 ng/ml TGF-β for 30 minutes with or without 10 μmol/L MG132, and immunoblots of whole-cell extracts for SnoN are shown. Subsequently, blots were reprobed for GAPDH to confirm approximately equal loading. A: Representative immunoblot of four experiments giving similar results. B: Densitometry of four independent experiments. C: HK-2 cells were transfected with Smad3 and Renilla control reporter constructs for 48 hours, before incubation with control medium or 1 ng/ml TGF-β for 6 hours with or without 10 μmol/L MG132. Data are presented as firefly/Renilla luciferase activity, normalized to control. D: HK-2 cells were incubated with control medium or 1 ng/ml TGF-β for 24 hours with or without 10 μmol/L MG132 before detection of PAI-1 mRNA by qRT-PCR. *P < 0.001 versus control; **P < 0.01 versus TGF-β. RLU, relative light units.