Follistatin-Like 1 Attenuates Ischemia/Reperfusion Injury in Cardiomyocytes via Regulation of Autophagy
Effect of FSTL1 pretreatment on the viability and apoptosis of H9C2 cells exposed to hypoxia/ischemia. (a) H9C2 cells were pretreated with FSTL1 at concentrations of 0, 5, 10, 20, and 40 ng/ml, and cell viability was determined by the CCK-8 assay after exposure to hypoxia/ischemia conditions for 24 h. Cells cultured under normoxic conditions were used as the control. The concentration of FSTL1 used for the following pretreatments was 10 ng/ml. (b) The CK-MB levels were determined with a commercial ELISA kit through three independent experiments. P < 0.05 vs. the control group, #P < 0.05 vs. the hypoxia/ischemia group. (c) Representative immunofluorescence images of H9C2 cells pretreated or not pretreated with FSTL1 under hypoxia/ischemia conditions. Cells cultured under normoxic conditions were used as the control. Proliferating cells were stained with EdU, and the total cells were stained with Hoechst 3344. (d) The percentage of EdU-positive cells among the total cells was calculated and analyzed. P < 0.05 vs. the control group, #P < 0.05 vs. the hypoxia/ischemia group. (e) Apoptosis as assessed by flow cytometry after Annexin V/PI staining. (f) Percentage of apoptotic cells. P < 0.05 vs. the control group, #P < 0.05 vs. the hypoxia/ischemia group. All experiments were repeated at least three times.
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