Fig. 4: Knockdown Wnt/β-catenin signaling in M2 macrophages abrogates protumor function of macrophages.

a, b BMDMs were transfected with β-catenin siRNA (S1 or S2) or control oligo (siR) for 24 h. The expression level of β-catenin was determined by qRT-PCR (a) and western blotting (b) (n = 3). c Knockdown of β-catenin in M2 BMDMs inhibited the ability of colony-forming ability of Hepa1-6 cells in the co-culture system. Representative images were taken at 14 days after incubation. The number of Hepa1-6 colonies was counted and compared (n = 3). d, e Knockdown of β-catenin in M2 BMDMs slowed down the wound-healing ability of Hepa1-6 cells in the co-culture system as determined by the scratch wound assay. The representative images of scratch wound healing were shown in (d). The migration distance of Hepa1-6 cells between two scratches was measured and calculated (e) (n = 3). f, g Knockdown of β-catenin in M2 BMDMs reduced the ability of migration (f, upper panel) and invasion (f, lower panel) of Hepa1-6 in the co-culture system as determined by a Transwell assay. The number of migration (g, left panel) and invasion (g, right panel) of Hepa1-6 cells was counted and compared (n = 3). h, i Knockdown of β-catenin was performed in differentially polarized macrophages that were irradiated with 20 Gy X-rays. Then, the irradiated cells were co-cultured with CFSE-labeled allogenic T cells for 5 days. The proliferation of T cells was determined by FACS (h) and was quantitatively compared among different groups (i) (n = 3). Bars, mean ± SD; *P < 0.05; **P < 0.01