Knockdown of Sestrin2 Increases Lipopolysaccharide-Induced Oxidative Stress, Apoptosis, and Fibrotic Reactions in H9c2 Cells and Heart Tissues of Mice via an AMPK-Dependent Mechanism

Figure 2

H9c2 cell viability is decreased by sesn2 knockdown upon LPS treatment. (a and b) Sesn2 knockdown H9c2 cells were incubated with LPS or LPS plus AICAR for 6 h, after which the ratio of Bax to Bcl-xL was determined by Western blotting. (c) Sesn2 knockdown cells were stimulated with LPS or LPS plus AICAR for 10 h, after which nuclei were observed by Hoechst staining (blue). (d) TUNEL-negative cells (only blue) and TUNEL-positive cells (blue plus green) were counted under a fluorescence microscope from the 4 fields of view randomly selected in each well, and then representative images were obtained for each group. (e) Caspase-3 activity in H9c2 cells was measured using a Caspase-3 Activity Assay Kit. (f) LDH release and (g) cell viabilities were calculated using a LDH assay reagent and cell viability assay reagent. (h) Sesn2 knockdown H9c2 cells were incubated with LPS or LPS plus NAC for 10 h, after which cell viabilities were measured. All graphs were obtained from more than three independent experiments. Data are presented as means; error bars represent ±SD (NS: not significant; ; ; ; ANOVA).

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