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Figure 2. Membrane recycling is impaired by the decrease of PtdIns[4,5]P2. Cells were labeled with FM 5–95 for 1 min at 37°C, washed with ice-cold Tyrode, warmed up to 37°C, and imaged for 60 min. Efflux kinetic curves of FM 5–95 following a 1 min pulse were obtained by quantification of fluorescent images. Fluorescent values were normalized to that of the first value at the beginning of the chase. Data points fit well to a double exponential decay (R2>0.99), giving a half-time of 
2 to 3 min for fast component and 20 to 25 min for slow component. Please note that after 60 min incubation, substantial amount of fluorescence was still retained in cells overexpressing 5-PPase. Data are mean ± SD; n = 4.
