Fig. 5: TINCR stabilized ATG5 mRNA by interacting with PTBP1. | Cell Death & Disease

Fig. 5: TINCR stabilized ATG5 mRNA by interacting with PTBP1.

From: The long noncoding RNA TINCR promotes self-renewal of human liver cancer stem cells through autophagy activation

Fig. 5

A, B Cellular fractionation and RNA-ISH assay were conducted to found cytoplasmic localization of TINCR. Scale bar, 100 mm. C catRAPID omics predicted the protein-RNA interactions. D, E RNA pull-down showed that PTBP1 interacted with TINCR. F RBPsuite (http://www.csbio.sjtu.edu.cn/bioinf/RBPsuite/) predicted the sequence motifs of PTBP1 binding sites. G RNAalifold (http://rna.tbi.univie.ac.at/cgi-bin/RNAWebSuite/RNAalifold.cgi) predicted the secondary structure of TINCR. The PTBP1 binding stem-loop structures in TINCR was indicated by red border. Pull-down assay and western blot were then performed to identify regions required for TINCR and PTBP1 interaction. H Relative ATG5 mRNA was detected by qRT-PCR when PTBP1 was overexpressed. I RIP assay was performed to detect the interaction between ATG5 and PTBP1 after TINCR knockdown. J Schematic description of the ATG5 mRNA 5′UTR, CDS, and 3′UTR. The interaction between PTBP1 and ATG5 mRNA 3′UTR was examined by pull-down analysis. K ATG5 mRNA stability was assessed in LCSCs with silencing TINCR or PTBP1 after being treated with actinomycin D. L ATG5 mRNA stability was evaluated in cells with overexpressed TINCR or combined with PTPB1 knockdown. Data are shown as means ± SD. *p < 0.05, **p < 0.01, ***p < 0.001.

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