Fig. 5: ERβ and WFDC3 attenuates CRC cell invasion through the suppression of TGFBR1.

a ERβ overexpression inhibits mRNA expression of TGFBR1, SMAD2, and SMAD3. b TGFBR1 overexpression rescues the ERβ-mediated reduction in SMAD2/3 protein expression levels. c Matrigel invasion assays of CRC cells co-transfected with Flag-TGFBR1 and Myc-ERβ or control plasmid. d Effect of SB525334 on ERβ depletion-induced SMAD2/3 downregulation in LoVo cells. e Effect of galunisertib on ERβ depletion-induced cell migration in LoVo cells. f WFDC3 overexpression inhibits mRNA expression of TGFBR1, SMAD2, and SMAD3. g TGFBR1 overexpression rescued the WFDC3-mediated decrease in SMAD2/3 protein expression levels. h Matrigel invasion assays of CRC cells co-transfected with Flag-TGFBR1 and Flag-WFDC3 plasmids. The bar graph indicates the number of invaded cells. i Effect of SB525334 on WFDC3 depletion-induced SMAD2/3 downregulation in LoVo cells. j Effect of galunisertib on WFDC3 knockdown-induced cell migration in LoVo cells. Bar graphs indicate the relative protein levels compared with β-actin or the number of migrated cells. Data are expressed as mean ± SD of at least three independent experiments. Statistics were performed using unpaired Student’s t-test, **P < 0.01, ***P < 0.001.