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. 2021 Sep 13;12:731980. doi: 10.3389/fphar.2021.731980

FIGURE 4.

FIGURE 4

Puerarin promoted SUMOylation as a potential cytoprotective mechanism against hypoxia/reoxygenation challenge. (A) Detection of LDH release from H9c2 cells. After 1 h treatment with puerarin-40 μM and ML-792-1μM, alone or in combination, H9c2 cells were exposed to hypoxia (0.1% O2) for 3 h and reoxygenation for 24 h. LDH release was measured by the LDH cytotoxicity assay kit. (B) Western blot analysis for COX2, galectin-3 and cleaved-PARP-1 expression. H9c2 cells were collected and analyzed by Western blotting with antibodies against COX2, galectin-3, cleaved-PARP-1. (C) Quantification of COX2, galectin-3 and cleaved-PARP-1 expression. The blots in panel B were quantified by a densitometric method. The results were shown as mean ± SD (n = 3). *p < 0.05. (D) Immunostaining of cellular 8-OHdG. H9c2 cells were probed with primary anti-8-OHdG antibody and visualized with Alexa Fluor 488-conjugated secondary antibody, whereas the cell nuclei were stained with DAPI. Scale bar, 50 μm. (E) Quantification of 8-OHdG formation. The results were presented as mean ± SD (n = 3). *p < 0.05.