Figure 2.

Bcr regulates actin stress fibers and focal adhesions in keratinocytes. (A–C) Control and Bcr KD NHEKs on coverslips were subjected to high calcium for 4 h, fixed, and processed to visualize F-actin using Alexa Fluor 488–conjugated phalloidin (A) or focal adhesions using an anti-vinculin antibody (C). Bar, 20 µm. (B) Representative images of cells with prominent, reduced, or no stress fibers are shown and quantified as described in Materials and methods. (D and E) Control and Bcr KD SCC9 cells were fixed and processed for immunofluorescence. F-actin was visualized using Alexa Fluor 488–conjugated phalloidin (D) and focal adhesions were visualized using an anti-phosphotyrosine antibody (PY99; E, inset). Bar, 20 µm. All images shown are representative of three independent experiments. KD of Bcr in SCC9 cells demonstrated a decrease in focal adhesions along the periphery of cells.