Figure 2: PtdIns5P is essential for Rac activation in NPM-ALK(+) cells.
From: Phosphatidylinositol 5-phosphate regulates invasion through binding and activation of Tiam1
(a) PtdIns5P mass levels in serum-starved Karpas 299 treated overnight or not with 800 nM of the PIKfyve inhibitor YM201636 or transduced with lentiviruses expressing shRNA directed against PIKfyve (sh1 and sh2). Data are shown as mean±s.e.m. (n=3 (Karpas 299 cells); n=3 (Cost cells); *P<0.05 using one sample t-test for the deviation from mean value of 100 in the control group). (b) Same experiment as in (a) with Cost cells. (c) Rac1 activation was monitored by pull-down from serum-starved Karpas 299 and Cost cells treated overnight or not with 800 nM YM201636. (d) Rac1 activation was monitored by pull-down from serum-starved Karpas 299 and Cost cells stably expressing shRNA directed against PIKfyve (sh1 and sh2). Data are shown as mean±s.e.m. (n=3; *P<0.05 using one sample t-test for the deviation from mean value of 1 in the control group). (e) Rac1 activation was monitored by pull-down from serum-starved Karpas 299 and Cost cells transduced by GFP-3xPHD wild type (WT) or GFP-3xPHD PI binding mutant (Mut)-expressing lentiviruses. Data are shown as mean±s.e.m. (n=3; *P<0.05 using one sample t-test for the deviation from mean value of 1 in the control group).
