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Fig. 5 | Journal of Experimental & Clinical Cancer Research

Fig. 5

From: LncGPR107 drives the self-renewal of liver tumor initiating cells and liver tumorigenesis through GPR107-dependent manner

Fig. 5

LncGPR107 recruited SRCAP to GPR107 promoter. a Biotin labeled lncGPR107 was generated through in vitro transcription, and incubated with oncosphere lysate for RNA pulldown, and the denoted band of lncGPR107 was identified as SRCAP by Mass spectrum. b The binding of lncGPR107 and SRCAP was confirmed by RNA pulldown and Western blot. c LncGPR107 truncates were generated (left panel) for RNA pulldown assay (right panel). d RNA electrophoretic mobility shift assay indicated the interaction of SRCAP and lncGPR107. The #8 region of lncGPR107 was used for RNA ESMA. e RNA immunoprecipitation (RIP) assay was performed with SRCAP antibody, and the enrichment of lncGPR107 was examined through realtime PCR. The oncospheres derived from HCC sample were used for RIP assay. f LncGPR107 and SRCAP subcellular location was examined through double fluorescence in situ hybridization (double-FISH) assay. Oncospheres were used for FISH, confirming the co-localization of SRCAP and lncGPR107 in oncospheres. Scale bars, 10 μm. g, h SRCAP ChIP assays were performed and GPR107 promoter enrichment was examined through realtime PCR. LncGPR107 depleted cells (g) and lncGPR107 overexpressed cells (h) were used for ChIP assays. Experiments were repeated at least three times

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