Fig. 4

Rho/ROCK signaling is required for stress fiber induction by FAM40 depletion. HUVECs were transfected with siRNAs targeting FAM40A, FAM40B or a control siRNA and seeded onto fibronectin-coated glass coverslips to form confluent monolayers. 72 h after transfection cells were treated with (a) 4 μg/ml C3 transferase for 2 h, or (b) 5 μM H1152 for 10 min after which they were fixed and stained for F-actin. Images are compressed stacks of 10–15 z-sections. Scale bars, 40 μm. Graphs show stress fiber content quantified from at least 150 cells per condition from 3 independent experiments. Error bars depict SEM values. *** (black) p < 0.001; Student’s t-test, compared to siControl; *** (grey) p < 0.001; Student’s t-test, comparison between siRNA+/− C3 transferase or H1152. (c) Intensity profiles are indicated for representative cells with the region scanned indicated as a red line in each image in panels 4A and 4B