Fig. 4
LncCCT4-2 in (HR + P)-EMVs is a potent mediator to inhibit oxidative stress and apoptosis in cardiomyocytes. (A) Heat map of differentially expressed microvesicular lncRNAs in (HR + D)-EMVs and (HR + P)-EMVs. (B) Analysis of differentially expressed lncRNAs in HR-EMVs, (HR + D)-EMVs and (HR + P)-EMVs by RT-qPCR. (C) HR-injured AC16 cells were co-cultured with HR-EMVs, (HR + D)-EMVs and (HR + P)-EMVs for 4 h, 12 and 24 h, respectively, and RT-qPCR was performed to detect expression of lncCCT4-2 in AC16 cells. (D) RT-qPCR analysis of lncCCT4-2 knockdown efficiency in sh-lncCCT4-2-(HR + P)-EMVs. HR-injured AC16 cells were co-cultured with HR-EMVs, (HR + D)-EMVs, (HR + P)-EMVs and sh-lncCCT4-2-(HR + P)-EMVs for 24 h, respectively: (E) Expression of lncCCT4-2 in AC16 cells was analyzed by RT-qPCR. (F) CCK-8 assay for AC16 cell activity. (G) Lactate dehydrogenase (LDH) activity of AC16 cell supernatants. (H) Flow cytometry detection of reactive oxygen species (ROS) levels in AC16 cells after DCFH-DA staining. (I) Quantitative analysis of ROS. (J) Flow cytometry detection of apoptotic rate in AC16 cells after Annexin V-FITC/PI double staining. (K) Quantitative analysis of apoptotic cells. (L-M) Western blotting analysis of Bax, Bcl-2, cleaved caspase-3 in AC16 cells, and the relative gray scales calculated by ImageJ software. These data were representative results (n = 3) of three repetitions. *P < 0.05, **P < 0.01, ***P < 0.001, ns, not significant. Data are expressed as mean ± SEM.