Cdc42 Null Cells Have Morphological Defects Involving the Formin FMNL2 and Focal Adhesions
All panels show EW7 Cdc42 f/f; ROSA26::Cre-ERT2; CDKN2−/− melanocytes treated with DMSO (control) or OHT (to delete Cdc42).
(A) Control and OHT-treated EW7 cells stained for endogenous FMNL2 and F-actin. White boxes denote zoom-in.
(B) EW7 cells expressing FMNL2 constructs as indicated.
(C) Length-to-width ratio.
(D and E) Proportion of cells in different shape categories (D) and roundness (E) was calculated; N = 31–37 cells per condition over three experiments; t tests compare data to the EW7 OHT GFP condition.
(F) Control and OHT-treated EW7 cells stained for vinculin and F-actin.
(G–I) Adhesion number per cell (G), adhesion area (H), and number of adhesions normalized to the cell area (I) were calculated using thresholded vinculin staining. N = 29–33 cells per condition over three experiments.
Graphs (C, E, G, H, and I) show mean ± SEM. ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001, t test with Welch’s correction. Scale bars, 10 μm. See also Figures S5 and S6.