Figure 4.
JPJD inhibited the EMT of CRC cells through TGF-β/Smad signaling pathway. (a) Western blot was applied to investigate the regulatory effect of JPJD on TGF-β/Smad signaling pathway. TGF-β-stimulated LoVo cells were treated with JPJD-M and/or TGFβRI/II inhibitor (LY2109761) for 48 hours. (b) Instantaneous inhibitory effect of JPJD on the activation of TGF-β/Smad signaling pathway. TGF-β-stimulated LoVo cells were treated with JPJD-M or LY2109761 only for 60 min or pretreated with LY2109761 followed by JPJD-M treatment for 60 min. (c) The process of EMT was detected followed by JPJD treatment and reexpression of Snail. (d) Real-time PCR was performed to measure the regulatory effect of JPJD on mRNA expression of E-cadherin and Snail. (e) Dual-luciferase assay for the inhibitory effect of JPJD on the promoter activity of CDH1 encoding E-cadherin Here, JPJD-L is 12.5 μg/mL, JPJD-M is 25 μg/mL, and JPJD-H is 50 μg/mL. ##P < 0.01, versus control LoVo cells; ∗P < 0.05 and ∗∗P < 0.01, versus only TGF-β-stimulated LoVo cells.