Figure 6. Vps60 and Snf7 polymers behave... | Rockefeller University Press

Figure 6.

$Vps60 and Snf7 polymers behave differently in vivo. Deletion of the VPS60 gene reduces the number of ILVs per endosome in yeast. (A–D) Two-dimensional cross sections and three-dimensional models from 200-nm-thick section electron tomograms of wildtype (A and B) and vps60∆ yeast cells (C and D). Endosomal limiting membranes are depicted in yellow. Scale bar: 100 nm. (E) Number of ILVs counted per endosome lumen in each strain. Wildtype ROI = 12, n = 7; vps60∆, ROI = 47, n = 4. P < 0.0001, two-tailed t test. (F) Western blots analyzing the distribution of Vps60-HA and Snf7 in detergent-solubilized membrane fractions resolved by rate-zonal density gradient centrifugation. Also indicated are the migrations of protein standards aldolase (158 kD), catalase (232 kD), and ferritin (440 kD). (G) Bar graphs representing the mean percentage of Vps60-HA and Snf7 in each gradient fraction examined in triplicate independent experiments; error bars indicate SD values. Source data are available for this figure: SourceData F6.$

Vps60 and Snf7 polymers behave differently in vivo . Deletion of the VPS60 gene reduces the number of ILVs per endosome in yeast. (A–D) Two-dimensional cross sections and three-dimensional models from 200-nm-thick section electron tomograms of wildtype (A and B) and vps60∆ yeast cells (C and D). Endosomal limiting membranes are depicted in yellow. Scale bar: 100 nm. (E) Number of ILVs counted per endosome lumen in each strain. Wildtype ROI = 12, n = 7; vps60∆, ROI = 47, n = 4. P < 0.0001, two-tailed t test. (F) Western blots analyzing the distribution of Vps60-HA and Snf7 in detergent-solubilized membrane fractions resolved by rate-zonal density gradient centrifugation. Also indicated are the migrations of protein standards aldolase (158 kD), catalase (232 kD), and ferritin (440 kD). (G) Bar graphs representing the mean percentage of Vps60-HA and Snf7 in each gradient fraction examined in triplicate independent experiments; error bars indicate SD values. Source data are available for this figure: SourceData F6.