Fig. 1.

Increased or decreased expression of Nup88 in HeLa cells affects cell migration. a Expression of endogenous Nup88, Nup98, and Nup214 in HeLa cells that stably overexpress GFP (lane 1) or Nup88-GFP (lane 2) was analyzed by immunoblotting. Unfilled, lightly shaded, and darker shaded arrowheads indicate GFP, endogenous Nup88, and Nup88-GFP, respectively. α-Tubulin was detected as a loading control. b Schematic outline of the scratch wound healing assay for Nup88-overexpressing HeLa cells. A confluent cell layer formed by HeLa cells overexpressing either GFP or Nup88-GFP was scratched to create a straight gap. After scratching, cells were further incubated with SFM for 24 h. c Cell migration from the initial gap (0 h) to the migration fronts (24 h) after scratching (scale bar 250 µm). A gap initially created in Nup88-overexpressing cells was completely closed within 24 h. d The average migration distance of cells 24 h after scratching was calculated. The average migration distance of GFP- and Nup88-GFP-overexpressing cells was 227 and 297 µm, respectively. Error bars indicate mean ± SD (n = 12); ***p < 0.001. e Schematic outline of the scratch wound healing assay for Nup88-knockdown HeLa cells. Parental HeLa cells transfected with siRNA were incubated with a serum-containing medium to form a confluent cell layer for 24 h, followed by incubation with SFM for 24 h until just before scratching. After scratching, cells were further incubated with SFM for 24 h. f Expression of Nup88 in HeLa cells transfected with control (siCONT) or NUP88-targeted (siNUP88) siRNA was analyzed by immunoblotting. α-Tubulin was detected as a loading control. g Cell migration from the initial gap (0 h) to the migration fronts (24 h) after scratching (scale bar 250 µm). h The average migration distance of cells 24 h after scratching was calculated. The average migration distance of siCONT- and siNUP88-transfected cells was 316 and 166 µm, respectively. Error bars indicate mean ± SD (n = 4); ***p < 0.001