Fig. 6
NEAT1 increases the mRNA stability of RPRD1B by recruiting hnRNPA2B1. A The datasheet shows part of NEAT1-interacting proteins obtained from CHART-MS. B GO analysis showed that the functions of NEAT1-interacting proteins were mostly enriched in RNA processing. C RIP assays show that NEAT1 is pulled down by hnRNPA2B1 antibody in RPRD1B-overexpressing HGC27 and SGC7901 cells. Immunoprecipitation with control IgG served as the negative control. (left and middle panels). Pull-down assays showed that hnRNPA2B1 was pulled down by NEAT1. Antisense of NEAT1 was used as negative control (right panel). D Positive correlation between RPRD1B expression and hnRNPA2B1 expression in TGCA data. E RIP-qPCR showing the enrichment of hnRNPA2B1 on the RPRD1B mRNA in RPRD1B-overexpressing HGC27 and SGC7901 cells. F Levels of the hnRNPA2B1 and RPRD1B proteins and mRNAs after RPRD1B inhibition in RPRD1B-overexpressing HGC27 cells. The results are summarized as the means ± SD of three independent experiments. G MeRIP-qPCR showing the enrichment of m6A in HGC27 cells after hnRNPA2B1 depletion. H The decay rate of the RPRD1B mRNA after treatment with 2.5 μM actinomycin D for the indicated times following hnRNPA2B1 knockdown in RPRD1B-overexpressing HGC27 cells. I RIP-qPCR showing the enrichment of hnRNPA2B1 on the RPRD1B mRNA in RPRD1B-overexpressing HGC27 cells with NEAT1 silencing. J The expression of RPRD1B was reduced following siNEAT1 transfection. K Representative IF staining images of the colocalization of NEAT1 and RPRD1B in RPRD1B-transfected HGC27 and SGC7901 cells. Scale bar, 20 μm. L, M Pull-down and RIP assay showing that NEAT1 interacted with RPRD1B in RPRD1B-transfected HGC27 and SGC7901 cells. Antisense of NEAT1 was used as negative control in pull-down assay. Immunoprecipitation with control IgG served as the negative control in RIP assay. GAPDH was served as the loading control. Data are presented as the means ± SD of three independent experiments. (*, P < 0.05, ***, P < 0.001)