Figure 5.

PM damage is followed by recruitment of ALG-2 and ESCRT-III leading to PM repair and shedding of the damaged membrane

(A) Micrograph of invasion pockets containing C. albicans. TR146 cells were transfected with CHMP2a-GFP (left), CHMP4B-mCherry (right). Side panels show the XY and YZ slices corresponding to areas marked by the dotted boxes.

(B) Micrograph of pockets containing wild-type or ece1Δ/Δ C. albicans. TR146 cells were transfected with CHMP2a-GFP (left), CHMP4B-mCherry (right), and PM bleb formation was assessed staining non-permeabilized cells with 10 μM FM4-64. Side panels show the XZ and YZ slices corresponding to the areas marked by the dotted boxes.

(C) Micrographs of TR146 cells treated with 30 μM Ece1-VIIa peptide or candidalysin. Cells were transfected with CHMP2a-GFP and PM bleb formation assessed by staining non-permeabilized cells with 10 μM FM4-64. (A–C) Outlines of TR146 cells indicated by dotted lines. Scale bars represent 5 μm.

(D) Confocal micrographs of TR146 cells infected with wild-type (left panel) or ece1Δ/Δ mutant C. albicans (right panel). ALG-2 was detected by immunofluorescence. Scale bar represents 10 μm.

(E) Quantification of ALG-2 accumulation as a function of invasion pocket length, from experiments like those in (D). Data are means ± SEM of ≥3 individual experiments.

(F and G) Micrographs of TR146 cells treated with 30 μM candidalysin. ALG-2 (F) and ALIX (G) detected by immunofluorescence and PM blebs detected staining the PM with ConA. Images are representative of ≥3 experiments of each type. See also Figure S2.