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Fig. 2 | Journal of Hematology & Oncology

Fig. 2

From: A cytoplasmic long noncoding RNA LINC00470 as a new AKT activator to mediate glioblastoma cell autophagy

Fig. 2

FUS interacted with LINC00470 and AKT to form a ternary complex in the cytoplasm. a The interaction of LINC00470 and FUS was detected through RIP assays in U251 cells. Data are presented as the mean ± S.E.M. of three independent experiments. **p < 0.01. b RNA pulldown showed binding between LINC00470 and FUS. c RIP assays showed that there was no interaction between LINC00470 and AKT in U251 cells. Data are presented as the mean ± S.E.M. of three independent experiments. d HEK293 cells were transfected with HA-AKT, Flag-FUS, and pcDNA3.1-LINC00470. Two-step co-immunoprecipitation verified their interaction. The expression levels of LINC00470, AKT, and FUS were measured with RT-qPCR and Western blotting, respectively. e The localization of AKT and FUS was detected by immunofluorescence staining in HEK293 cells. f The co-localization of AKT and FUS was detected by immunofluorescence staining in U251 cells. g Left, the interactions between endogenous FUS and AKT in the cytoplasm and nucleus were measured by co-immunoprecipitation; right, an RNA pulldown assay showed binding between endogenous LINC00470 and AKT in the cytoplasm and nucleus of U251 cells transfected by si-FUS. h Western blotting detected the expression of FUS in GBM cells transfected by si-FUS. Expression levels of AKT and p-AKTS473 were measured by Western blotting in GBM cells that re-expressed LINC00470 in FUS-KD GBM cells. i Western blotting detected the expression levels of p-AKTS473 in the cytoplasm and nucleus of U251 cells transfected by si-LINC00470

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