FIGURE 8:

Depletion of ATP9A does not affect EGF trafficking. (A) HeLa cells were transfected with siRNAs against LacZ, ATP9A-1, or ATP9A-2; serum starved for 3 h; incubated at 4°C for 60 min with Alexa Fluor 488–conjugated EGF; washed; and incubated at 37°C for the indicated times. Cells were fixed and stained for EEA1. Scale bars: 20 μm. (B) Pixel intensity of Alexa Fluor 488–conjugated EGF was estimated at the indicated times using MetaMorph software. Data are shown as the ratio of the mean of cellular EGF intensities between cells depleted of ATP9A and control cells at each time point. Graph is representative of three independent experiments, and 100–200 cells of each sample were analyzed. Graphs show means ± SE.