Fig. 3: Nf2 deletion resulted in defective cell migration, adhesion, and attachment.

a A scratch-wound assay was performed to examine MSC migration that isolated from Nf2 control and Nf2 mutant mice. The healing distances were calculated at each time point and analyzed (n = 8) (b). c Primary osteoblasts were plating on fibronectin precoated plates with no fetal bovine serum for different time points, and the non-attached cells were washed, and the remaining osteoblasts were stained by crystal violet and counted for analysis, and Nf2 knockout led to significantly diminished cell adhesion. d The cultured adherent Nf2 mutant osteoblasts was significantly reduced after cell detachment experiment using 0.2% EDTA for 10 min. e F-actin staining by Phalloidin and β-tubulin staining showed that cytoskeletal assembly was disorganized in Nf2 mutant osteoblasts. f Rhotekin-RBD beads were added to lysate to pull down the RhoA, and Western Blotting were performed to measure the activity of RhoA in Nf2 mutants (BK030, Cytoskeleton, Inc.) and the data showed that RhoA activity was significantly decreased in Nf2 mutant MSC. Scale bar= 50 μm in (a, e). Data were expressed as means ± SD and each dot represents an individual biological replicate. P values were calculated by unpaired Student’s t-test with two-tailed analysis without adjustments. *p < 0.05. **p < 0.01, ***p < 0.001.