Fig. 1.
Vav2 and Vav3 are required to maintain epithelial traits in breast cancer cells. a Defects shown by indicated 4T1 cell lines on primary tumorigenesis and lung metastasis according to earlier work [26]. The epithelial and mesenchymal phenotypes scored in the current work are also included. b, c Representative example of the morphology of indicated 4T1 cell lines in 2D (b) and 3D (c) cultures (n = 3 independent experiments). Scale bars, 50 μm. KD2/3 (A) and KD2/3 (B) are two independent clones of Vav2;Vav3 knockdown 4T1 cells [26] (Table S1). d Representative immunoblot showing the abundance of indicated endogenous proteins in lysates from 4T1 cells lines shown on top (n = 3 independent experiments). The primary antibody used in the immunoblot is shown in the right. For loading control, we used the abundance of endogenous actin. Asterisks mark nonspecific bands. e Representative immunofluorescence analysis showing the abundance and subcellular localization of endogenous E-cadherin (green color), rhodamine phalloidin-stained F-actin (red color), and β-catenin (green color) in indicated 4T1 cell lines (left) (n = 3 independent experiments). Scale bar, 25 μm