Figure 4.

Integrin-activation dependent Sept-7 regulation enhances sensitivity of cells to changes in ECM cues and contributes to their ECM remodeling function

(A) Representative wide-field fluorescence images of MEFs transfected with non-targeting or Sept-7 siRNA plated on soft (0.4 kPa) or stiff (60 kPa) polyacrylamide gels and probed for F-actin; scale bar, 20 μm.

(B) Quantification of cell area of NT or Sept-7 KD MEFs on soft or stiff polyacrylamide gels, n = 3 replicates, 59–61 cells.

(C) Wide-field fluorescence images showing NT and Sept-7 KD MEFs stained for F-actin and plated on 10 μg/mL FN, 0.1 μg/mL FN, and 0.1 μg/ml FN + MnCl²; scale bar, 20 μm.

(D) Quantification of cell area for NT and Sept-7 KD MEFs plated on 10 μg/mL FN, 0.1 μg/mL FN, and 0.1 μg/ml FN + MnCl², n 2–3 replicates, 29–59 cells.

(E) Wide-field fluorescence images showing NT and Sept-7 KD MEFs plated for 8 h on 10 μg/mL FN and co-stained with F-actin (cyan), and FN (yellow); scale bar, 100 μm.

(F) Quantification of the average area of FN clearance per cell in NT and Sept-7 KD cells, 2 replicates, n = 555–645 images. All statistics performed with Kruskal-Wallis and Dunn’s multiple comparisons test or Mann-Whitney test, ∗∗∗∗p < 0.0001, ∗∗∗p < 0.001, ∗∗p < 0.01, ∗p < 0.05, ns, not significant, orange horizonal lines show medians.