Research Paper Volume 12, Issue 21 pp 22291—22312

Figure 6. PVT1 regulates BCLAF1 expression via sponging miR-194-5p. The relative expression patterns of BCLAF1 were detected in paired bladder carcinomas tissues and normal tissues (A). The relative expression of BCLAF1 was reduced by si-PVT1 (B). Overexpressing miR-194-5p down-regulated BCLAF1 expression and knockdown of miR-194-5p up-regulated BCLAF1 expression in BC cells (C). Dual-luciferase reporter assay showed BCLAF1-Wt and miR-194-5p mimics co-transfection restrained luciferase activity (D). Immunoprecipitation assays were used to identify proteins associated with BCLAF1. The anti- BCLAF1 and IgG antibody were incubated with cell extracts and CDC20 was identified to be a BCLAF1-binding protein in the cancer cell line (E). Knockdown of PVT1 decreased BCLAF1 etc. expression in BC cells (F). Overexpressing miR-194-5p decreased BCLAF1 expression and knockdown of miR-194-5p increased BCLAF1 expression in BC cells (G). Knockdown of PVT1 decreased MPP2 and MPP9 expression in BC cells (H). Overexpressing miR-194-5p decreased MPP2 and MPP9 expression and knockdown of miR-194-5p increased MPP2 and MPP9 expression in BC cells (I). Knockdown of BCLAF1 reversed BCLAF1 expression promotion induced by overexpression PVT1 in BC cells (J). IP pyrolysis products are used for proteomics detection (Ka–f): a-KEGG, b-GO, c-COG, d-IPR, e-function annotation, f-subcellular localization (*P < 0.05, **P < 0.01).