Fig. 2
IMP2 promotes the stability of ZFAS1 by directly binding to the m6A site of ZFAS1. a Schematic diagram of IMP2 directly binding to ZFAS1 to improve ZFAS1 stability. b The expression of ZFAS1 after overexpression or silencing IMP2 in HCT116 and SW620 cells by qPCR. c Rescue experiments detecting the ZFAS1 expression levels after co-transfected with IMP2 silencing and ZFAS1 overexpression or IMP2 overexpression and ZFAS1 silencing vectors in HCT116 and SW620 cells assayed by the qPCR assay. d Reducing lncRNA half-life (t1/2) of ZFAS1 decay after treated with shIMP2 vectors in HCT116 and SW620 cells. e The heat map illustrating the differentially expressed lncRNAs in HEK293T. f CLIP database showing the motif of ZFAS1 targeting IMP2 and the motif of m6A site predicted by online software (http://www.starbase.sysu.edu.cn). g Cuilab (http://www.cuilab.cn/) online platform showing qualified m6A binding sites (RGGAC) of ZFAS1. h Bioinformatics online software predicting the specific binding sequence and sites of ZFAS1 secondary structure and IMP2 protein (http://www.tartaglialab.com/). i MOE docking platform showing the specific docking sites between ZFAS1 tertiary structure and IMP2 protein. j Co-localization of ZFAS1 and IMP2 protein detected by the combination of ISH and IF assays in HCT116 and SW620 cells. Scale bar = 5 μm. k RIP followed by qPCR showed binding of the ZFAS1 with anti-m6A in HEK293T cells. l The relative expression of ZFAS1 after treated with IMP2-WT and IMP2-Mut (KH3-4) vectors in HCT116 and SW620 cells. m RNA pull-down followed by western blot showed in vitro binding of the ZFAS1-Wild and ZFAS1-Mutant probes with IMP2 protein after IMP2 silencing in HEK293T cells. n Reducing lncRNA half-life (t1/2) of ZFAS1 decay after treated with ZFAS1-Mut vectors in HCT116 and SW620 cells. Data were shown as mean ± s.d. of at least three independent experiments. *P < 0.05; **P < 0.01; *** P < 0.001; **** P < 0.0001 and ns no significance