Fig. 4

Vimentin and actomysoin are implicated in TNT formation after RASSF1A depletion. (a) Quantification and (b) representative images of Vimentin expression in HBEC-3 cells transfected with siNEG or siRASSF1A. (c) Quantification and (d) representative images of Vimentin expression in A549 cells transfected with pcDNA control or RASSF1A. (e) Quantification and (f) representative images of TNT formation in HBEC-3 cells transfected with either siVimentin alone or in combination with siRASSF1A. (g) Representative images (i and ii are zoom of the corresponding arrowheads for respectively DMSO, Paclitaxel, Nocodazole and Blebbistatin treatment) and (h) quantification of TNT number in control and RASSF1A depleted HBEC-3 cells after incubation with paclitaxel (10 nM), nocodazole (10 μM) or blebbistatin (5 μM) for 24 h before fixation and staining with α-tubulin. The TNTs are show with arrowheads. Roman numerals mark the examples of the TNT in the zoomed images. (c) The mRNA expression was assayed using RT2 ProfilerTM Cell motility PCR Array (Qiagen). β₂-microglobulin was used as an internal control. Values are the mean ± SEM of three independent experiments in almost 200 cells. Statistical significance was calculated and p value are indicated by asterisks: *p < 0.05, **p < 0.01, *** < 0.001