Fig. 5: LncRNA-6195 combining with protein ENO1. | Cell Death & Disease

Fig. 5: LncRNA-6195 combining with protein ENO1.

From: A novel lncRNA, TCONS_00006195, represses hepatocellular carcinoma progression by inhibiting enzymatic activity of ENO1

Fig. 5

a Silver-stained SDS-PAGE gel-containing proteins derived from RNA pulldown by lncRNA-6195 and negative control RNA. The red arrow indicates the gel cutting for mass spectrometric analysis. b Western blot analysis of ENO1 derived from RNA pulldown by lncRNA-6195, its antisense RNA and negative control RNA. Input is the total protein used for RNA pulldown. c RT-PCR analysis of RNAs derived from RIP assays in HepG2 cells. d Agarose electrophoresis of the PCR products. e Western blot analysis of ENO1 derived from RNA pulldown by negative control RNA, lncRNA-6195, its antisense RNA, and different fragments. f Graphic illustration of ENO1 and different truncation mutants(RCPA: 97–237aa, region required for repression of c-myc promoter activity; SB: 370–373aa, region required for substrate binding; IP: 405–434aa, region required for interaction with plasminogen). g Western blot analysis of Flag-tagged ENO1 and its mutants. h RIP experiments were performed using Flag antibody on extracts from HepG2 cells transfected with Flag-tagged full-length or mutant ENO1 expression vectors. The purified RNAs were analyzed by RT-PCR. i Agarose electrophoresis of the PCR products. (***P < 0.001. Student’s t test. Data are presented as mean ± SD.)

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