Fig. 3

β-escin downregulates HER2, p95HER2, HER3 and Akt expression. A–D Western blots for HER2, p95HER2, phospho-HER2 (Y1221/1222), phospho-p95HER2, HER3, phospho-HER3 (Y1289), Akt, and phospho-Akt expression in BT474 (A, C) and JIMT-1 cells (B, D) following exposure to β-escin (10-20 μM and 20-30 μM, respectively) for 48 h. GAPDH was used as an internal loading control. The ratio of the protein content is represented in the right panels (*p<0.05). (E, F) Immunofluorescence analysis of HER2 (red) with DAPI (nuclei, blue) in BT474 (E) and JIMT-1 cells (F) after treatment with β-escin (20 and 30 µM, respectively) for 24 h. Intensity profiles represent HER2 expression with green signal fluorescence. G Immunoblot analyses of HER2 and p95HER2 protein content in HER2- and p95HER2-overexpressing MDA-MB-231, and SKBR3 cells. (H, I) Immunofluorescence analyses of HER2 (green) and ICD-HER2 (green, 4B5) with DAPI (nucleus, blue) in HER2- and p95HER2-overexpressing MDA-MB-231 cells in the presence of β-escin (20 and 30 µM, respectively) for 24 h. Intensity profiles were analyzed with the histogram tool in the Zen blue software and the horizontal line (white dotted line) indicates 100 intensity units (range of y-axis, 0-250 units)