Figure 3.

ACP KD affects adhesion formation. HGFs were transfected with control siRNA and ACP siRNA and plated on FN for 30 min in the presence of IL-1 and coimmunostained for ACP and paxillin or talin. A) Confocal microscopy images of immunofluorescence localization of ACP (red) and paxillin (green) in cells spreading on FN. Higher magnification of leading edge of cells is shown in boxed regions. Lysates of control siRNA and ACP siRNA–transfected cells were immunoblotted for ACP to confirm ACP KD. GAPDH was used as a loading control. In the lower left panel, the histogram displays the mean ± sd number of FAs in the cell periphery (<5 μm from the cell membrane) that were immunostained with paxillin; n = 10–12 cells/group. The histogram in the lower left panel shows the means ± sd length of FAs immunostained with paxillin in cell periphery (<5 μm from the cell membrane; n = 10–12 cells/group). FAs were quantified using ImageJ. Differences between groups were measured by ANOVA and post hoc Tukey’s test. B) Identical procedures were conducted for ACP KD and immunostaining for talin, followed by analysis of talin-stained FAs including computation of the means ± sd number of peripheral FAs (left panel) and the means ± sd FA length; n =10–12 cells/group. Differences between groups were measured by ANOVA and post hoc Tukey’s test.