Journal: PLoS Genetics

Article Title: Endocytic recycling via the TGN underlies the polarized hyphal mode of life

doi: 10.1371/journal.pgen.1007291

Figure Lengend Snippet: (A) Scheme of the effectors subordinated to RabB RAB5 in early endosomes (EEs)(see text). (B) Normal localization of ChsB in a rabBΔ mutant. (C) Normal localization of ChsB in a vps33 ts strain at 28°C and following a shift to 42°C. (D) Left, delocalization of ChsB in rabCΔ ; Middle, plot of average intensities of ChsB in wt and rabCΔ apical domes determined from 2 x 50 pixel arch-shaped linescans as in the scheme. P values estimated with an unpaired t -test; Right plot, linescans (mean values ± S.E.M. bars) of maximal intensities across the whole width of the same tips used for the plots. (E) Delocalization of ChsB from the apical dome in the vps52Δ mutant; middle and right plots and statistical analysis as in (D). Note that for (D) and (E), hyphal tip images were not deconvolved to better display the differences between the wt and the mutants.

Article Snippet: To estimate the amount of polarized GFP-ChsB in wt, rabCΔ and vps52Δ strains, middle planes of z-stacks were used to determine fluorescence intensities along 50 px-long and 2 px wide ROIs covering the plasma membrane in the apical domes (with Metamorph).

Techniques: Mutagenesis

Journal: PLoS Genetics

Article Title: Endocytic recycling via the TGN underlies the polarized hyphal mode of life

doi: 10.1371/journal.pgen.1007291

Figure Lengend Snippet: Cells were stained with the vital dye CMAC (7-amino-4-chloromethylcoumarin) to reveal late endosomes and vacuoles (CMAC shown in magenta in color composites). The main images are MIPs of unprocessed z-stacks, but the insets were deconvolved to remove apical haze. Note that the contrast of the 28°C vps52 Δ image has been adjusted to reveal the cytosolic haze, with ‘empty holes’ corresponding to the nuclei. Colocalization of ChsB with CMAC in the 37°C vps52 Δ sample was complete.

Article Snippet: To estimate the amount of polarized GFP-ChsB in wt, rabCΔ and vps52Δ strains, middle planes of z-stacks were used to determine fluorescence intensities along 50 px-long and 2 px wide ROIs covering the plasma membrane in the apical domes (with Metamorph).

Techniques: Staining