Fig. 2.

The tumor microenvironment induces the de-differentiation of bASCs. A-C bASCs (ln-aT, ln-dT, ob-aT and ob-dT) were stained for αSMA, FSP1 and CD29 for FACS analyses. CAFs (ln-CAF and ob-CAF) were isolated and stained as positive controls. Quantification of αSMA (A), FPS1 (B) and CD29 (C) are shown as bar graphs. The results are from three independent experiments (n = 3, 60.000 cells for each condition and in each group) and presented as mean ± SEM. D Representative images of bASCs and CAFs stained for αSMA (green), phalloidin (red) and DNA (DAPI, blue) are shown. Red boxes indicate measured areas. Scale: 25 μm. Inset scale: 12.5 μm. E The evaluation of the mean fluorescence intensity of αSMA is presented as scatter plots. The results are from three independent experiments (n = 3, 90 cells for each condition and in each group) and presented as mean ± SEM. F Cellular extracts from bASCs were prepared for WB analysis with antibodies against αSMA, COL1A1, caveolin-1, AKT, pAKT and pGSK3β. GAPDH and β-actin served as loading controls. G Quantification of the αSMA signal in WB is shown, relative to the corresponding amount of GAPDH. The results are from three independent experiments and presented as mean ± SEM. H and I Relative gene levels of ACTA2, TAGLN and CTGF, important myCAF marker genes (H), and relative gene expression of CSF3, CXCL10, IL1β and HAS1, iCAF marker genes (I), are shown for bASC subgroups (ln-dT, ln-aT, ob-dT and ob-aT). The results are from three independent experiments and presented as bar graphs with mean ± SEM. Student’s t test was used in (A-C) and (G-I). Unpaired Mann-Whitney U test was used in (E). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. ∗p < 0.05, ∗∗∗p < 0.001