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. 2020 May 29;9(6):1355. doi: 10.3390/cells9061355

Figure 2.

Figure 2

R87C mutation in CYFIP enhances lamellipodium protrusion at low Rac GTPase levels. (A) Western blotting of distinct cell lines to probe for expression levels of endogenous CYFIP and/or Rac GTPases. (B) Quantification of lamellipodia formation in B16-F1 CYFIP1/2+Rac1/2/3 KO cells (clone #3/4), harboring low Rac expression and transfected with indicated EGFP-tagged CYFIP1 constructs, as described in Figure 1A. Statistical significance was assessed by two-sample, two-sided t-test. n.s.: not statistically significant; ** p < 0.01; *** p < 0.001 (C) Live cell imaging of CYFIP1/2+Rac1/2/3 KO cells (clone #3/4) expressing EGFP-tagged CYFIP1 constructs. Upper panels show phase contrast images, and bottom panels EGFP fluorescence. Scale bar, 10 µm. (D) Quantification of cell edge advancement. n gives the number of cells analyzed. Box and whisker plots represent data as follows: boxes correspond to 50% of all data points (25–75%), and whiskers to 80% (10–90%). Lines and numbers above boxes correspond to medians. Statistical significance was assessed by non-parametric, Mann-Whitney-Rank-Sum test, n.s.: not statistically significant; *** p < 0.001. Bottom part shows representative kymographs.