Fig. 5: MTR4 regulates AS of its target pre-mRNA by recruiting PTBP1.
From: MTR4 drives liver tumorigenesis by promoting cancer metabolic switch through alternative splicing
a AS of WT and two MTR4 binding motif mutant (MT) including poly(A) MT in GLUT1 minigene in PLC/PRF5 cells. Data are presented as mean value ± s.d. Two-tailed, unpaired t-test. p value is indicated. n = 3 independent experiments. b AS of WT and two MTR4 binding motif mutant (MT) including poly(A) MT in PKM minigene in PLC/PRF5 cells. Data are presented as mean value ± s.d. One-way ANOVA with a Dunnett’s multiple comparison test. p value is indicated. n.s., non-significant. n = 3 independent experiments. c Impact of PTBP1 knockdown on AS of PKM minigene in PLC/PRF/5 cells with or without MTR4 overexpression. The levels of PKM1 and PKM2 isoforms were analyzed using primers described in (b). Representative data from two independent experiments are shown. d The co-immunoprecipitation analysis of the endogenous MTR4, hnRNPA1, and PTBP1 in PLC/PRF/5 cells in the absence or presence of RNase. The cell lysate was subjected to immunoprecipitation (IP) with indicated antibodies. The presence of various proteins in the immunoprecipitate was examined by western blotting using indicated antibodies. Because IP grade anti-MTR4 antibody is not commercially available, we did not perform the MTR4 IP. Representative data from two independent experiments are shown. e RIP analysis of the binding of PTBP1 to the WT and MTR4 binding motif mutant PKM minigene in PLC/PRF5 cells. Data are presented as mean values. n = 2 independent experiments. Source data are provided as a Source Data file.
