PIAS1 Alleviates Hepatic Ischemia-Reperfusion Injury in Mice through a Mechanism Involving NFATc1 SUMOylation

Figure 2

The effect of PIAS1 on NFATc1 SUMOylation and the inflammatory response and apoptosis of hepatocytes. (a) The mRNA expression of PIAS1 and NFATc1 in the liver tissues of sham-operated and HIRI mice determined by RT-qPCR (). (b) The protein expression of PIAS1 and NFATc1 in the liver tissues of sham-operated and HIRI mice determined by western blot analysis (). (c) The SUMOylation of NFATc1 in 293 T cells transduced with Flag-NFATc1 and HA-SUMO1 detected by Co-IP assay. (d) The SUMOylation of endogenous NFATc1 in AML12 cells detected by Co-IP assay. (e) The SUMOylation of NFATc1 in H/R-exposed AML12 cells treated with oe-PIAS1 or combined with ML-792 detected by Co-IP assay. (f) Apoptosis of H/R-exposed AML12 cells detected by TUNEL staining ( vs. control, # vs. H/R, & vs. H/R + oe-PIAS1). (g) RT-qPCR detection of PIAS1 and NFATc1 expression in H/R-exposed AML12 cells treated with oe-PIAS1 or combined with oe-NFATc1. (h) The mRNA expression of TNF-α, IL-1β, IL-6, Bax, Bcl2, and caspase-3 in H/R-exposed AML12 cells treated with oe-PIAS1 or combined with oe-NFATc1 measured by RT-qPCR. (i) The protein expression of TNF-α, IL-1β, IL-6, Bax, Bcl2, and cleaved caspase-3 in H/R-exposed AML12 cells treated with oe-PIAS1 or combined with oe-NFATc1 measured by Western blot analysis. (j) Apoptosis of H/R-exposed AML12 cells following treatment with oe-PIAS1 or combined with oe-NFATc1 detected by TUNEL staining. In (g)–(j), vs. oe-NC + sh-NC, # vs. oe-PIAS1 + oe-NC. The cell experiment was repeated 3 times.

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