Fig. 5

FMNL2 is required for invasion in A375 and WM266.4 melanoma cells. a A375 and WM266.4 melanoma cells were transfected with control or FMNL2-specific siRNA duplexes. Control and knockdown cells were then assayed for their ability to invade in vitro in an inverted Boyden chamber assay (Kitzing et al. [6]). FMNL2 knockdown inhibited invasion in both A375 and WM266.4 cells. b Equivalent samples of A375 cell lysates from (a) were subjected to SDS-PAGE and immunoblotted to detect FMNL2 and FMNL3. The pan-FMNL2 blot was stripped and re-probed with anti α-tubulin as a loading control. c Equivalent samples of WM266.4 cell lysates from (a) were subjected to SDS-PAGE and immunoblotted to detect FMNL2. The pan-FMNL2 blot was stripped and re-probed with anti α-tubulin as a loading control. d A375 cells were transfected with control and FMNL2-targeted siRNA duplexes and then infected with control lentiviral vector or lentiviral vectors expressing the indicated untagged full-length FMNL2 isoforms. The cells were then assayed for their ability to invade as in (a). Images show a representative Z-stack projection from the invasion assays. e Equivalent samples of cell lysates from (d) were subjected to SDS-PAGE and immunoblotted to detect FMNL2. α-tubulin was used as a loading control. f Quantification of data shown in (d). N = 3, error bars = SEM. *indicates P < 0.05 where noted. There was no significant difference between TQS and ITM or TQS and YHY