Fig. 6

circFOXP1 promotes PKLR expression by interacting with PTBP1 in GBC cells. a left, the protein level of PTBP1 was detected using western blot analysis in GBC tissues compared with adjacent normal tissues. Right, immunohistochemical staining and mRNA expression of PKLR in GBC tissues compared with adjacent normal tissues. Arrow, PKLR expression was shown. b RIP assays with PTBP1 antibody were performed to detect PKLR enrichment in NOZ and SGC-996 cells. All data are shown as mean ± S.E.M., n = 3. c A RBP map was used to analyze the binding sites between PTBP1 and the 3’UTR and CDS region of PKLR. PTBP1 was pulled down by the 3’UTR and CDS wt RNA probe but not by the mut RNA probe in NOZ and SGC-996 cells. d, e The mRNA enrichment of PKLR was determined using RIP-qRT-PCR after silencing of circFOXP1 in NOZ and SGC-996 cells or overexpression of circFOXP1 in GBC-SD cells. All data are shown as mean ± S.E.M., n = 3, **P < 0.01. f The expression levels of circFOXP1 were determined using western blot analysis after transfection of NOZ and SGC-996 cells with sh-NC, sh-circFOXP1–1 or sh-circFOXP1–2 or GBC-SD cells with pLCDH-vector, pLCDH-circFOXP1, pLCDH-circFOXP1+ si-PTBP1, n = 3 or more