Fig. 4
From: A subset of activated fibroblasts is associated with distant relapse in early luminal breast cancer
CDH11 expression by CAF-S1 enhances cancer cell migratory properties. a CDH11 mRNA levels in CAF subsets (CAF-S1 to S4) from BC of Curie cohort described in [22]. Data expressed in log2+1 are from RNAseq and shown as mean ± SEM. b Representative views of CDH11 immunostaining in stromal compartment in controls (left) and cases (right) (scale bar = 50 μm). c CDH11 H score in CAF according to recurrence status (N = 102, 51 controls, 51 cases). p value is from Wilcoxon test. d, e Number of MCF7 (d) and T47D (e) cancer cells alone or after 3 days of co-culture with CAF-S1 transfected with non-targeting siRNA (CAF-S1 + siCTL) or with CDH11-targeting siRNA (N = 3, each in triplicates), after normalization on carboxylate beads. f, g Same as in d, e for percentage of alive MCF7 (f) and T47D (g) cancer cells assessed by FACS analysis (DAPI−) (N = 3 in triplicates). h–j Migration of MCF7 (d) and T47D (e) cancer cells (evaluated by Transwell assays) without (h–j) or with (k–m) contact with CAF-S1 transfected with control or CDH11-targeting siRNA. h Representative images of lower-side Transwell using MCF7 (left) and T47D (right) into the well and CAF-S1 transfected with control or CDH11-targeting siRNA. i, j Number of cancer cells (MCF7 (i) and T47D (j)) that have migrated in Transwell lower-side. Data are means ± SEM (N = 4). Schemas show Transwell designs used, with two different siCDH11 tested (siCDH11#1 and siCDH11#2). (K-M) Same as in h–j with CAF-S1 cultured in contact of cancer cells. In k, arrowheads indicate cancer cells and arrows CAF-S1, distinguished by morphological criteria. (N = 4). n Images of E-cadherin (red) and DAPI (blue) staining (top) or of E-cadherin (red), F-actin (green), and DAPI (blue) co-staining (bottom) in BC cells alone or in presence of CAF-S1 transfected with control or CDH11-targeting siRNA. Scale bars, 20 μm. o Quantification of E-cadherin staining per BC cell area (at least six images analyzed per condition). Data are mean ± SEM) (n = 3). a. u., arbitrary units. p values from Mann-Whitney test