Figure 5. PI(3)P is recruited to endosomes concomitantly with Rab5.

HeLa cells, stably expressing mApple-Rab5 and transiently transfected with the PI(3)P marker, GFP-FYVE, were treated for 20 min with nigericin, washed and imaged over 3 hr, as described in Figure 4A. (A) Time-lapse images of a representative endosome to show transient Rab5 recruitment accompanied by PI(3)P. A time-lapse video of the endosome at 1 min interval is available in Figure 5—video 1. (B) Corresponding graph of normalised mean fluorescence intensity of Rab5 and FYVE at the rim of the endosome in (A) over the time the endosome was detectable. (C) Averaged Rab5 and PI(3)P kinetics of 19 endosomes. Error bars represent standard deviation. Representative graph of three independent experiments. Numerical data for all analyzed endosomes is available in Figure 5—source data 1.

Figure 5—figure supplement 1. PI(3)P is recruited to endosomes concomitantly with Rab5.

HeLa cells, stably expressing mApple-Rab5 and transiently transfected with the PI(3)P marker, GFP-FYVE, were treated for 20 min with nigericin, washed and imaged over 3 hr, as described in Figure 4A. (A) Time-lapse images of a representative endosome to show transient Rab5 recruitment accompanied by PI(3)P. (B) Corresponding graph of normalised mean fluorescence intensity of Rab5 and FYVE at the rim of the endosome in (A) over the time the endosome was detectable. (C) Averaged Rab5 and PI(3)P kinetics of 16 endosomes. Error bars represent standard deviation. (D) Averaged Rab5 and PI(3)P kinetics of 15 endosomes. Error bars represent standard deviation. (C) and (D) represent two independent experiments of a total of three.

Figure 5—video 1. Representative endosome to show transient Rab5 recruitment accompanied by PI(3)P.

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