Fig. 7

RanBPM inhibits HDAC6 mediated cell migration. a Control or RanBPM shRNA HEK293 cells transfected with either empty vector (EV), WT-RanBPM (WT) or Δ360 RanBPM (Δ360) were harvested 30 h before the scratch or 24 h after the scratch. Whole cell extracts were analyzed by western blot and hybridized with the antibodies indicated. b Control shRNA HEK293 cells were treated with either DMSO or 2 μM Tubacin for the indicated time points. Whole cell extracts were analyzed by western blot and hybridized with the antibodies indicated. c Confluent monolayers of control or RanBPM shRNA HEK293 cells were transfected with either EV, WT or Δ360 RanBPM and were cultured in the presence of 2 mM hydroxyurea for 24 h, and pretreated with either DMSO (black bars) or 2 μM Tubacin (white bars) for 4 h. Cells were then scratched and width of the wound was assessed at the time of the scratch and 24 h later. Fold migration was calculated by normalizing the average wound width at 24 h to the average wound width at 0 h and each sample was normalized to control shRNA EV transfected, DMSO treated samples. Results are averaged from three independent experiments, with error bars indicating SEM. P < 0.05 (*). d Representative images of Control or RanBPM shRNA HEK cells transfected and treated as above following the scratch (0 h) and 24 h later