Figure 5.

shFXN hBMVEC are deficient in total polymerized actin and lack peripheral actin. (A) EVEC and (B) shFXN hBMVEC are incubated with Phalloidin-Texas Red and Hoechst, (C) the ratio taken to quantify total filamentous actin per cell, then normalized to EVEC controls. (D) 6.4 µm of 2–5 membranes per cell were quantified for F-actin (gray value) and averaged into a histogram. The membrane interface (peak) is represented by the dotted line. 300nm surrounding the peak was quantified as the cortical actin region (CAR), as represented by the shaded region. Individual datapoints of the cortical ROI are represented in the inset with significance and the %deficit in F-actin. (E) Values and %difference with statistics for the peak membrane F-actin. Student’s T-test α =0.05; ns = not significant, * p < 0.05, ** p < 0.01. Total phalloidin analysis (C): EVEC; n = 30, shFXN; n = 27. Membranes analyzed (D,E): EVEC; n = 191, shFXN; n = 222. The images in panels A and B were cropped from their original size while maintaining the scale of the scale bar.