Fig. 4.

Endosomal trafficking to vacuoles was greatly compromised in vps41 pollen tubes. (A–H) Endosomal trafficking to vacuoles revealed by a 2S albumin–mRFP fusion protein marker. Fluorescent and bright-field images of wild-type and vps41 pollen tubes are shown. (A and B) Fluorescent and bright-field images of wild-type pollen tubes at 1.5 hag. (C and D) Fluorescent and bright-field images of vps41 pollen tubes at 1.5 hag. (E and F) Fluorescent and bright-field images of wild-type pollen tubes at 3.5 hag. (G and H) Fluorescent and bright-field images of vps41 pollen tubes at 3.5 hag. At least 20 independent pollen tubes were observed for each genotype and each time point. (Scale bar, 5 μm.) (I–M) FM4-64 uptake assays. Intracellular accumulation of FM4-64 is shown within 10 min after incubation in 5 μM FM4-64 in a wild-type pollen tube (I) and vps41 pollen tube (J). (K) Statistical quantification of relative FM4-64 uptake within 10 min. At least 50 independent pollen tubes were used for statistical analysis for each genotype. Error bars indicate SD. Distribution of FM4-64 is shown after 240 min of incubation in 5 μM FM4-64 in a wild-type pollen tube (L) and a vps41 pollen tube (M). (N) Statistical quantification of relative FM4-64 transport to tonoplast at 240 min; 20 independent pollen tubes were used for statistical analysis for vps41 pollen tubes, and 14 independent pollen tubes were used for statistical analysis for wild-type pollen tubes. (Scale bar, 5 μm.)