Fig. 5: miR-21 regulates the autophagy of colorectal cancer cells through PTEN/AKT/TFEB. | Cell Death & Disease

Fig. 5: miR-21 regulates the autophagy of colorectal cancer cells through PTEN/AKT/TFEB.

From: An autoregulatory feedback loop of miR-21/VMP1 is responsible for the abnormal expression of miR-21 in colorectal cancer cells

Fig. 5

a Autophagic structures (AV, AL, and IM) were observed in parental RKO and RKO-miR-21-KO cells by transmission electron microscope. Number of autophagic structures was counted. b RKO and RKO-miR-21-KO were treated with glucose starvation or bafilomycin A1, the LC3 (LC3I and LC3II) expression was measured by Western blot. c The protein expression level of PTEN, phosphorylated AKT (Ser473), VMP1, LC3, and nuclear TFEB were measured by Western blot in RKO and RKO-miR-21-KO cells. d RKO and RKO-miR-21-KO were treated with VO-Ohpic at concentrations of 1–4µM for 24 h, and immunoblot analysis of phosphorylated AKT (Ser473), VMP1, LC3 expression and the protein amount of nuclear TFEB. e RKO and RKO-miR-21-KO were treated with Afuresertib at concentrations of 1–4 µM for 24 h, and immunoblot analysis of VMP1, LC3 expression, and the protein amount of nuclear TFEB. f The protein expression level of VMP1, LC3, total TFEB, and the protein amount of nuclear TFEB were measured by Western blot after knock-down of TFEB expression by siRNA in RKO and RKO-miR-21-KO cells. (Scale bars, 0.5 µm) Data are presented as mean ± SD. **P < 0.01.

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