Fig. 3

Features of fibroblasts exhibited by CAAs. A Adipocytes were cocultured with melanoma cells ( lines: WM1341D, A375, WM9 and Hs294T) present on Transwell inserts and then obtained CAAs as well as the non-differentiated 3T3L1 fibroblasts and control adipocytes were seeded on coverslips coated with gelatin-FITC (green). After 16 h of incubation, cells were fixed and stained using phalloidin-Alexa Fluor 568 to visualize F-actin (red). Areas of gelatin degradation are visible as dark holes on a green background. White arrows indicate stress fibers. Scale bar: 25 μm. B Western blot analysis of protein level of vimentin and TGFβ receptor III (TGFβRIII) in cell lysates of 3T3L1 cells; control adipocytes, and adipocytes cultured with melanoma cells growing on Transwell inserts. The signal was normalized to total protein content assessed by Ponceau S staining. The mean of at least three biological repetitions ± SD is shown. Asterisks indicate statistically significant differences between control adipocytes, 3T3L1 fibroblasts and CAAs. The significance levels were set at p ≤ 0.05 (*), p ≤ 0.01 (**), or p ≤ 0.0001 (****).