Figure 4.

CAIF overexpression increased miR‐16 expression by increasing demethylation. AC16 cells were transfected with the CAIF expression vector or miR‐16 mimic, and overexpression was confirmed by RT‐qPCR at 48 h posttransfection (A). The effects of CAIF overexpression on miR‐16 (B) and the effects of miR‐16 overexpression on CAIF (C) were analyzed by qRT‐PCR. Methylation‐specific PCR was performed to analyze the effects of CAIF overexpression on miR‐16 gene methylation (D). The experiments were repeated in triplicate, and the mean ± SD values were compared. C, control cells without transfection; CAIF, cardiac autophagy inhibitory factor; M, methylated PCR product; miR, microRNA; NC, cells transfected with empty vector or NC miRNA; qRT‐PCR, quantitative reverse‐transcription polymerase chain reaction; U, unmethylated PCR product. *p < .05