Figure 6
From: CCAR1 5′ UTR as a natural miRancer of miR-1254 overrides tamoxifen resistance
Mechanistic characterization of miR-1254 and CCAR1 5′ UTR in modulating cellular sensitivity to tamoxifen. (A) miR-1254 targets predicted by TargetScan and miRanda with further GO analysis. NCOA1, NCOA3, EGFR, ERBB2 and SNAI1 are selected. (B) Relative luciferase activity of wild-type or mutant 3′ UTRs with forced expression of miR-1254. Error bars indicate SEM. ***P< 0.001, N.S., not significant. (C) Western blot analysis of CCAR1- and miR-1254-targeted genes upon forced expression of miR-1254 in MCF-7 TAM-R cells or miR-1254 antagonism in MCF-7 cells for 48h. (D-E) Relative luciferase activity of 3′ UTR luciferase reporter (D), mRNA levels (E) of miR-1254-targeted genes in MCF-7 TAM-R cells transfected with CEG or vector. Error bars indicate SEM. ***P< 0.001. (F-G) Relative luciferase activity of 3′ UTR luciferase reporter (F) and mRNA levels (G) of miR-1254-targeted genes in MCF-7 TAM-R cells transfected with shNS or shCCAR1. Error bars indicate SEM. ***P< 0.001. (H) Protein levels of indicated genes upon forced expression of CEG in MCF-7 TAM-R cells or depletion of CCAR1 in MCF-7 cells. Student's t-test. See also Supplementary information, Figure S7.
