Fig. 4: Mechanism dissection of how DHT/circRNA-BCL2 induces autophagic cell death in EnzR PCa cells: via regulating the AMBRA1 expression. | Cell Death Discovery

Fig. 4: Mechanism dissection of how DHT/circRNA-BCL2 induces autophagic cell death in EnzR PCa cells: via regulating the AMBRA1 expression.

From: High-dose-androgen-induced autophagic cell death to suppress the Enzalutamide-resistant prostate cancer growth via altering the circRNA-BCL2/miRNA-198/AMBRA1 signaling

Fig. 4

A, B RNase R assay to determine the sensitivity of circBCL2 by RNase R digestion in EnzR1-C4-2 (A) and EnzR4-C4-2B (B) cell lines. C Venn diagram for prediction of the potential miRNAs using two miRNA related websites (miRDB and Regrna2.0). D Seven miRNA candidates can physically interact with circRNA-BCL2 and showed up after screening EnzR-C4-2 cells by biotin-circRNA-BCL2 pull-down assay. E The qRT-PCR was performed to show quantification of circRNA-BCL2 related miRNAs expression in EnzR-C4-2 cells after 50 nM DHT treatment. F Argonaute2 (AGO2) IP assay to detect 14 candidate genes mRNA in AGO2 complex and results revealed that 8 genes have less miRNAs binding in EnzR-C4-2 cells after 50 nM DHT treatment. G, H Western blot assays were performed to show candidates genes expression in EnzR-C4-2 cells after 50 nM DHT treatment. I, J Western blot assays show AMBRA1, LC3, and p62 expression after oe-circRNA-BCL2 in EnzR1-C4-2 (I) and EnzR4-C4-2B (J) cell line with 50 nM DHT. K, L Western blotting show AMBRA1, LC3 and p62 expression after sh-AMBRA1 in EnzR1-C4-2 (K) and EnzR4-C4-2B (L) cell line with 50 nM DHT. M, N Colony formation assays show cell growth with 50n M DHT when sh-AMBRA1 is transduced in EnzR1-C4-2 (M) and EnzR4-C4-2B (N) cell lines. Data are presented as means ± SD. *p < 0.05, **p < 0.01, NS not significant.

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