Fig. 4

STAT3/FoxQ1 contributes to TAMs-induced EMT and macrophages attraction. (a) Western blot analysis of CRC cells (HCT116 and HT29) alone, IL6-supplemented CRC cells, TAMs-co-cultured CRC cells, and IL6 depleted TAMs-co-cultured CRC cells. (b) Western blot analysis of HCT116 cells alone, IL6-supplemented HCT116, TAMs-co-cultured HCT116 in the presence or absence of Stattic (15 μM). (c) Relative expression levels of representative EMT related factors were detected in HCT116 with or without 48 h of TAMs co-culture as determined by RT-PCR. Error bars, SEM. (d) Western blot of FoxQ1 from whole-cell lysates extracted from CRC cells (HCT116 and HT29) co-cultured with TAMs for the indicated times. (e) Western blot of colorectal cancer cells (HCT116 and HT29) transfected with FoxQ1 expression vector or empty vector and indicated siRNAs (si-STAT3) and incubated with IL6 for 48 h afterwards. (f) Western blot of CRC cells (HCT116 and HT29) transfected with STAT3 expression vector or empty vector and indicated siRNAs (si-FoxQ1). (g) CCL2 mRNA expression in CRC cells (HCT116 and HT29) with or without 48 h of TAMs co-culture; Error bars, SD. (h) ELISA assay of CCL2 protein secretion of CRC cells (HCT116 and HT29) transfected with FoxQ1 expression vector or empty vector. Error bars, SD. (i) THP-1 cells migration towards CRC cells (HCT116 and HT29) alone and TAMs-co-cultured CRC cells with or without anti-CCL2 Ab; Error bars, SD. ***P < 0.001