Figure 3.

LINC00174 modulates the I/R-induced myocardial injury by downregulating p53

(A and B) Mouse primary myocardial cells were pretreated as indicated before H/R treatment.

(A) The relative expression of LINC00174 in mouse primary myocardial cells after hypoxia treatment for 4 h was measured by qRT-PCR; n = 3.

(B) The expression of p53 in mouse primary myocardial cells after H/R treatment was measured by western blotting. β-Actin was used as the loading control.

(C) Negative control shRNA (shNC) or LINC00174 shRNA (shLINC00174) was transfected into mouse primary aortic endothelial cells; 48 h later, the exosomes in supernatant were harvested to determine the relative expression level of LINC00174 by qRT-PCR; n = 3.

(D) Mouse primary myocardial cells were pretreated as indicated before H/R treatment. The expression of p53 in mouse primary myocardial cells was measured by western blotting after H/R treatment for 4 h. β-Actin was used as the loading control.

(E–G) Mouse primary myocardial cells were pretreated as indicated before H/R treatment.

(E) The cell apoptosis after H/R treatment was determined by TUNEL assay; cell nuclei were stained by DAPI. Scale bar, 200 μm.

(F) The formation of autophagosomes was assessed by immunofluorescence staining; cell nucleus was stained with DAPI. Scale bar, 10 μm.

(G) The expression of autophagy-related proteins was measured by western blotting. β-Actin was used as the loading control.

(A–G) The data represented 1 of 3 independent experiments. (A and C) Data were represented as means ± SDs. p values were determined by 1-way ANOVA, followed by Tukey post hoc test. ∗p < 0.05 and ∗∗p < 0.01.