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. 2020 Feb 8;14(4):808–828. doi: 10.1002/1878-0261.12634

Figure 5.

Figure 5

PCAT7 disrupts miR‐324‐5p‐mediated suppression on TGFBR1. (A) The predicted miR‐324‐5p binding sites in TGFBR1 (TGFBR1‐Wt) and the designed mutant sequence (TGFBR1‐Mut) were indicated. (B) Luciferase reporter assay for the luciferase activity of TGFBR1‐3′‐UTR‐Wt and TGFBR1‐3′‐UTR‐Mut reporters in the presence of miR‐324‐5p mimics or miR‐324‐5p inhibitor in PCa cells. Data are shown as mean ± SD. *P < 0.05. (C) Western blot analysis showed miR‐324‐5p mimics decreased, while miR‐324‐5p inhibitor dramatically increased TGFBR1 protein level in PCa cells. (D) Luciferase reporter assay showed the impact of miR‐324‐5p mimics and TGFBR1 on the luciferase activity of TGF‐β signaling. Data are shown as mean ± SD. *P < 0.05. (E, F) Transwell assay (E) and Matrigel invasion assay (F) to show the migration and invasion ability of PCa cells in the indicated groups. Data are shown as mean ± SD. *P < 0.05. (G) Luciferase reporter assay of TGFBR1‐3′‐UTR‐Wt and TGFBR1‐3′‐UTR‐Mut reporters in PCa cells with the expression of PCAT7 and miR‐324‐5p changed. Data are shown as mean ± SD. *P < 0.05. (H) Western blot analysis for TGFBR1 in PCa cells with the expression of PCAT7 and miR‐324‐5p changed. (I) Ago2 RNA immunoprecipitation (RIP) assay for the amount of PCAT7 in the indicated group. Data are shown as mean ± SD. *P < 0.05.