Fig. 5: ALG2 promotes ANXA11 condensation to increase the lipid order of lysosomes in cells.
From: ANXA11 biomolecular condensates facilitate protein-lipid phase coupling on lysosomal membranes
a U2OS cells under hypotonic conditions (95% ddH2O, 5% DMEM, pH 7.0) expressing a fluorescently labelled lysosome marker (TMEM192-Halo-JF647) and ANXA11 (ANXA11-mEm). The cytosolic pool of ANXA11 was photobleached to reveal ANXA11-membrane associations. The white ROI indicates the displayed zoomed regions. Scale bar - 5 µm. b A representative image of a hypotonic U2OS cell with fluorescently labelled lysosomes (TMEM192-Halo-JF647) incubated in 100 nM PK dye to extract the relative order of lysosomal membrane lipids. The white ROI within the zoomed panels indicates an example analysis segment. Scale bars - 10 µm (left panel) and 1 µm(right panel). c Quantitation of the relative order (φ) of lysosomal membranes as displayed in (b) from U2OS cells alone, or expressing either ANXA11, ANXA11 + ALG2, or ANXA11 + CALC. Mean ± SD. One-way ANOVA with Tukey’s multiple comparison, **p = 0.0021, n = 3 repeats (82–146 lysosomes).
