Fig. 4

H19 binds to hnRNPA2B1 specially. a Coomassie blue staining of gel after SDS-PAGE separated the proteins which were immunoprecipitated with full length of H19 and its antisense RNA in HCT116 cells by RNA pull-down assays. The arrow shows the position of hnRNPA2B1. b Western blot analyses following RNA pull-down assays in HCT116 cells confirmed the interaction between H19 and hnRNPA2B1. Input, total proteins. Pull down, proteins immunoprecipitated by RNA. c RIP assay followed by qRT-PCR suggested H19 binds to hnRNPAA2B1. d qRT-PCR to analyze the subcellular distribution of H19 after compartmentalization of cytoplasmic and nuclear fractions in HCT116 and DLD1 cells. β-actin and U6 act as cytoplasmic and nuclear controls, respectively. e Western blot to analyze total level of hnRNPA2B1 between H19 overexpressed and control cells. f Western blot analyzed the effect of H19 on the protein level of hnRNPA2B1 in nuclear and cytoplasm. g Western blot analyzed the protein level of hnRNPA2B1 in nuclear and cytoplasm of H19 depleted and control cells. h Immunofluorescence were performed to investigate the subcellular localization of hnRNPA2B1 of H19 overexpression and control cells. Scales bars = 10um Student’s t-test. *P < 0.05, **P < 0.01, ***P < 0.001