Fig. 2

FSCN1 inactivation in H295R cells causes defects in spreading after plating and proliferation. (A) Western blot showing expression of FSCN1, SF-1 and GAPDH in control #1 and #2 and in FSCN1 KO #1 and #2 H295R clones in basal conditions and after treatment with Dox (1 μg/ml) for 72 h. (B) Staining of control and FSCN1 KO cells with phalloidin (actin cytoskeleton; in red) and paxillin (focal adhesions; in green). Scale bar, 5 μm. (C) FSCN1 KO cells have a spreading defect after plating. The histogram shows the percentage of spread cells 48 h after plating for control and FSCN1 KO H295R cells. Data are derived from the combined analysis of control #1 and #2 (orange) and FSCN1 KO #1 and #2 H295R clones (violet). Data for individual clones are shown in Figure S1. n (independent experiments) = 6. Mean ± SD is shown. ****P < 0.0001, t test. Bottom: Representative micrographs of control and FSCN1 KO cells taken 48 h after plating. Scale bar, 20 μm. (D) FSCN1 KO cells proliferation is slower compared with control cells. Data are derived from the combined analysis of control #1 and #2 (orange line) and FSCN1 KO #1 and #2 H295R clones (violet line). Cells were cultured without adding Dox in the culture medium. Data for individual clones are shown in Figure S3. n (independent experiments) = 6. Mean ± SD is shown. ***P < .001, t test.