Figure S5.
Panel A: The image shows immunofluorescence staining for galectin3 in wild-type (WT) and VRAC knockout (KO) bone marrow-derived macrophages (BMDMs) under different conditions. The conditions include isotonic media (Iso), hypo-osmotic media (Hypo), and treatment with the lysosomal permeabilisation agent LLOME. The images are labeled as WT and KO for each condition. Galectin3 puncta positive cells are highlighted with white arrows. The scale bar is 25 micrometers. Panel B: A line graph depicting the relative fluorescence of DQ-BSA over time in WT and VRAC KO BMDMs under different conditions. The conditions include isotonic media (Iso), hypo-osmotic media (Hypo), and treatment with bafilomycin A (BafA). The x-axis represents time in hours, and the y-axis represents DQ-BSA relative fluorescence. The graph includes data points for WT Iso, KO Iso, WT Hypo, KO Hypo, WT BafA, and KO BafA. The values shown are the mean with standard error of the mean (SEM).
Lysosomal integrity and function are unaffected by hypo-osmotic media and loss of VRAC. (A and B) Immunofluorescence for galectin-3 in WT and VRAC KO BMDMs incubated in iso- or hypo-osmotic media, or with the lysosomal permeabilization agent LLOME (1 mM) for 5 h (n = 3). Galectin-3 puncta–positive cells are highlighted with white arrows (scale = 25 µm) (B) Time-lapse of DQ-BSA fluorescence in WT and VRAC KO BMDMs incubated in iso- or hypo-osmotic media (n = 4). BafA1 (100 nM) was included as a control of reduced lysosomal function. Values shown are the mean ± the SEM.