Skip to main content
. 2023 May 9;42(25):2074–2087. doi: 10.1038/s41388-023-02703-9

Fig. 3. Vimentin is required for in vitro cancer cell migration and invasion.

Fig. 3

A A scratch wound assay was used to evaluate cell migration. Representative images are shown at 0 and 6 h following scratch formation. Wound area closure was compared to the starting value and quantified for KPV+/+ (n = 11) and KPV−/− (n = 17) cells; each point represents a separate scratch wound. B Cell invasion through a Matrigel-coated transwell was measured over 48 h. Invasive index is the mean number of cells invaded per 20× magnification imaging field. KPV+/+ (n = 9) and KPV−/− (n = 11) cell invasion data are plotted so that each point represents data from a single transwell assay. C KPV+/+ and KPV−/− spheroids were suspended in type I collagen and spheroid growth was tracked over 48 h. Spheroid area was quantified relative to the initial area of each spheroid (n = 4 independent experiments). Scale bar: 200 µM. D KPV+/+ cells were treated with withaferin A (WFA; 5 or 10 µM) or DMSO vehicle control for 6 h. Cells were stained for vimentin (white; dotted line represents cell outline). Scale bar: 10 µm; inset: 5 µm. E KPV+/+ cells were treated with vehicle (n = 6) or 5 µM WFA (n = 4) and were subjected to a scratch wound assay. Wound area was quantified at 6 h. F KPV+/+ cells were plated atop a Matrigel-coated transwell and were treated with vehicle control (n = 6) or 5 µM WFA (n = 9); invasion was quantified at 48 h via invasive index as described above. Data are presented as the mean ± standard deviation. The p-values were calculated using an unpaired, two-tailed t-test, except for panel C, in which data were compared using a repeated-measure two-way ANOVA with multiple comparisons. (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001).