Extended Data Fig. 7: Localization of LRRC8A in BMDMs.
From: Proton-gated anion transport governs macropinosome shrinkage
(a) Lrrc8a3xHA/3xHA mice54 that express a LRRC8 protein C-terminally tagged with 3 copies of the HA-epitope from the native genomic locus were used to determine the subcellular localization of the essential VRAC subunit LRRC8A. Anti-HA antibodies detected the protein in the plasma membrane of Lrrc8a3xHA/3xHA BMDMs but not in WT control macrophages. (b) LRRC8A is present at the plasma membrane of Lrrc8a3xHA/3xHA BMDMs. There is no significant co-localization with endolysosomal markers rab5 (early endosomes), rab7 (late endosomes) or LAMP1 (lysosomes). Pearson’s R calculated from 18 cells (Rab5), 20 cells (Rab7), 16 cells (LAMP1). (c) 4 min after M-CSF addition, LRRC8A can be detected in early macropinosomes (co-stained by rab5) in ≈5-10% of cells, but not in rab7-positive mature macropinosomes. All small panels represent magnified macropinosomes or regions of interest indicated by white arrowheads. Pearson’s R calculated from 15 cells (Rab5, 4min), 10 cells (Rab7, 4min), 5 cells (Rab5, 7min), 10 cells (Rab7, 7min). Mean ± s.e.m. Scale bars: 10 µm and 1 µm for the magnification panels. Source numerical data are available in source data.
