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. 2017 Oct 30;114(46):E9903–E9912. doi: 10.1073/pnas.1716009114

Fig. 4.

Fig. 4.

Specific engagement of VIM by FiVe1 induces defects in chromosomal alignment during metaphase. (A) Western blotting analysis of phosphorylated VIM protein content from FOXC2-HMLER cells treated for 24 h with FiVe1 (500 nM). Quantification (B) and representative images (C) of immunofluorescent staining for P-S56-VIM from FOXC2-HMLER cells treated with the indicated doses of FiVe1 (n = 3, mean and SD). (D) Quantification of multinucleated, FLAG positive FOXC2-HMLER or HEK293T cells 72 h after transfection with the indicated VIM expression constructs (n = 3, mean and SD). (E) Representative images of FOXC2-HMLER cell immunostained for FLAG protein content 72 h after transfection with the indicated VIM expression constructs. (Scale bar, 10 μm.) (F) Relative viability measurements of FOXC2-HMLER cells 7 d after transduction with lentiviruses encoding the indicated overexpressed transgene products (n = 8, mean and SD). (G) Representative confocal images of thymidine-synced FOXC2-HMLER cells at metaphase immunostained for VIM and β-tubulin (TUB) (FiVe1, 250 nM). (Scale bar, 10 μm.) (**P < 0.005, ***P < 0.0005; NS, not significant; t test).