Figure 2.

Effects of DN rab4, DN rab11, and a PI3K inhibitor on LacCer recycling. (A) Normal HSFs or NPFs were cotransfected with the DsRed2-Nuc plasmid and either DN rab4 or DN rab11 constructs. After 48 h, the recycling assay was performed as in Figure 1. Transfected cells were identified on the basis of red fluorescent protein in the nucleus. T, transfected cell; UT, untransfected cell. Bar, 15 μm. (B and C) Fluorescence intensity remaining in the cells after 10 or 40 min of recycling was quantified by image analysis and expressed as a percentage of fluorescence present at 0 min. Values are mean ± SD (n = 60 cells; three independent experiments). The role of PI3Ks on lipid recycling from the EEs was studied using the PI3K inhibitor WM.