The modulation of PI(3,4,5)P₃ levels affects macropinosome formation.

<p>A: HEK-Flp-In cell monolayers were treated with either 65 µM LY294002 or carrier (0.2% Ethanol) for 30 minutes at 37°C prior to pulsing with dextran-TR as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0013763#s4" target="_blank">Materials and Methods</a> in the continued presence of the drug or carrier. The mean number of macropinosomes/100 cells was quantitated as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0013763#s4" target="_blank">Materials and Methods</a>. Error bars represent S.E.M. B: HEK-Flp-In cell monolayers transfected with pEGFP-Grp1-PH for 24 hours were imaged using the 100× oil immersion objective of an Olympus IX-81 OBS Real Time microscope before and after treatment with 65 µM LY294002. Time following LY294002 treatment is indicated in bottom right hand corner of each panel. C: HEK-Flp-In cells transiently transfected with pIRES2-EGFP, pIRES2-EGFP-PTEN, or pIRES-EGFP-PTEN(G129E) were co-transfected with pmCherry, pmCherry-SNX9, or pmCherry-SNX18 and assayed for macropinosome formation as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0013763#s4" target="_blank">Materials and Methods</a>. The mean number of macropinosomes/100 transfected cells was quantitated over 3 replicates of 500 transfected cells for each condition. Error bars denote S.E.M.