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Fig. 7 | Journal of Experimental & Clinical Cancer Research

Fig. 7

From: LncRNA CASC9 interacts with CPSF3 to regulate TGF-β signaling in colorectal cancer

Fig. 7

CASC9 and CPSF3 coregulate the expression of TGFβ2. a RT-qPCR of selected genes related to the ranked pathways and tumorigenesis in CPSF3-knockdown HCT-116 cells. b, c Correlation of CASC9 and TGFβ2 (b) and of CPSF3 and TGFβ2 (c) expression in 155 human CRC tissues. Expression data for CASC9, CPSF3, and TGFβ2 were downloaded from the lncRNAtor database. d The decay of TGFβ2 mRNA was monitored by RT-qPCR in CASC9- or CPSF3-knockdown HCT-116 cells by blocking mRNA synthesis using actinomycin D (ActD, 5 μg/mL) for the indicated time points upon normalization to RNA input levels. e RIP revealed that CPSF3 binds TGFβ2 mRNA in HCT-116 cells. Validation of CPSF3-immunoprecipitation efficiency by western blotting. f The mRNA levels of SMAD7, TGFβ2, and TERT were determined in HCT-116 cells transfected with pcDNA3.1-CASC9–202. g The mRNA levels of TGFβ2 were determined in HCT-116 cells cotransfected with pcDNA3.1-CASC9–202 and siCPSF3. h The TGFβ2 protein and phosphorylated SMAD3 levels were determined in CASC9-knockdown or -overexpressing HCT-116 cells. i The TGFβ2 protein and phosphorylated SMAD3 levels were determined in CPSF3-knockdown CRC cells. The data are presented as the mean ± s.d. *P < 0.05, **P < 0.01 by Student’s t-test (a, e, f, g)

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