Fig. 4

Residual tumor cells are surrounded by extensive stroma exhibiting a fibrotic response after BGJ398 treatment. a Mammary glands after 10-day BGJ398 treatment as compared to controls. b Histological analysis of tumors in BGJ398 treatment groups (10 days of treatment) as compared to control group treated with vehicle. Upper panel: Hematoxylin and eosin (H&E) staining. Lower panel: Trichrome staining. c Quantification of percentage of collagen in tumors. **p < 0.005, comparisons represent control versus 10 days after BGJ398 treatment. Values are shown as mean ± SEM. d Immunofluorescence double staining of proliferation marker Ki67 (green) in HA (red)-positive tumor cells and apoptosis marker cc3 (red) in tumor cells after 10 days BGJ398 treatment as compared to control group treated with vehicle. e Quantification of Ki67-positive cells in HA-positive cells after 10 days BGJ398 treatment as compared to a control group treated with vehicle alone, ****p < 0.0001; quantification of cc3-positive cells after 10 days BGJ398 treatment as compared to control group treated with vehicle alone, ***p = 0.0009. f Immunofluorescence double staining of K5 and K8 in tumors treated with BGJ398 for 10 days as compared to tumors in control group treated with vehicle. Upper row: K8 (green) and K5 (red), magnification 200×. Lower row: K8 (red) and K5 (green), magnification 400×. g Immunofluorescence staining of α-SMA, tenascin-C, CD31 and S100A8 in tumors treated with BGJ398 for 10 days as compared to tumors in control group treated with vehicle. h Quantification of vasculature after 10 days BGJ398 treatment as compared to control group treated with vehicle alone, ****p < 0.0001. i Quantification of S100A8-positive cells after 10 days BGJ398 treatment as compared to control group treated with vehicle alone, **p = 0.0012. α-SMA alpha smooth muscle actin, cc3 Cleaved caspase 3, DAPI 4′, 6-Diamidino-2-phenylindole