Fig. 5

PTEN dependent anterior plasma membrane translocation of PH-Crac in EF. a-b PHCrac-GFP was redistributed asymmetrically to the leading edge of the electrotaxing WT cells. EF = 30 V/cm. c Latrunculin A treatment did not affect the EF-triggered asymmetric redistribution of PH-Crac in WT cells. d PH-Crac anterior relocalization was abolished in pten⁻ cells, while evenly distributed global membrane translocation remained. e Time-lapse analysis of fluorescence intensity ratio comparing membrane translocation of PTEN-GFP (posterior vs anterior) and PHCrac-GFP (anterior vs posterior) plasma membrane in EF. The asymmetric relocalization of PHCrac-GFP was fully abolished in pten⁻ cells. f Relative fluorescence intensity analysis further confirmed that the EF-triggered anterior redistribution of PHCrac-GFP was not affected by latrunculin A in WT cells, and significantly increased membrane translocation was recorded at both anterior and posterior of pten⁻ cells. g Membrane fraction cell lysates from the cells treated with the indicated duration of EFs were analyzed by immunoblotting with antibodies against PHCrac-GFP in WT or pten⁻ cells, respectively