Fig. 4

WNT4 promotes the recruitment and activation of fibroblasts. a and b The expression of infiltrating α-SMA-positive cancer associated fibroblasts (CAFs) in different groups: LoVo-Scramble or LoVo-shWNT4 (n = 7), SW480-WNT4-vector or SW480-WNT4-HA (n = 5). Scale bar, 200 μm. CAFs, red; DAPI, bule. c and d Typical heterospheroids formed in different groups: LoVo-Scramble or LoVo-shWNT4, SW480-WNT4- vector or SW480-WNT4- HA. CAF, red; CRC cells, green; Scale bar, 20 μm. e Stronger gel contraction was observed after the NF#1 and NF#2 treatment with WNT4 (400 ng/mL). f and g Increased expression of fibronectin (FN) and α-SMA were observed by immunofluorescence. CAFs and NFs treated with TGF-β (10 ng/mL) served as positive controls. f fibronectin-red, phalloidin-green, scale bar, 50 μm. g α-SMA-red, scale bar, 20 μm. h Increased expression of FN and α-SMA were observed by western blotting analysis. A two-tailed Student’s t test was used for statistical analyses. Measurement data were presented as the mean ± SD. *P < 0.05