Figure 6.

miR-200 s and/or their targets Fli-1 and TCF12 are involved in the expression of ECM-associated genes in CAFs. (a) Heat map illustrating the hierarchical clustering of altered ECM-associated genes in CAFs and paired NFs. (b) qRT-PCR analysis of FN and LOX expression in 15 paired CAF and NF samples that were freshly isolated from breast tumor tissues. U6 was used as an internal control. The data are shown as the normalized fold change in CAFs compared with NFs. (c and d) Western blot analysis of FN and LOX expression in the indicated CAFs or NFs. (e) mRNA expression of FN and LOX as determined by qRT-PCR in the indicated CAFs or NFs. β-Actin was used as an internal control. The data are presented as the mean±S.D. (n=3; *P<0.05, Student's t-test). (f) Western blot analysis of FN and LOX expression in CAF-sh/TCF12 or NF/TCF12 cells. (g) LOX protein expression was analyzed by western blot after re-expressing Fli-1 in CAF/miR-200c cells or knocking down Fli-1 in NF-sh/miR-200c cells. (h) FN protein expression was evaluated by western blot after re-expressing TCF12 in CAF/miR-141 cells or knocking down TCF12 in NF-sh/miR-141 cells