Figure 4.
Panel A shows GCaMP6s fluorescence trace showing cytosolic calcium changes of WT macropahges after LLOMe and thapsigargin treatment over time in the presence of calcium in the extracellular media. Panel B shows the calcium response of macrophages in calcium-free media to thapsigargin followed by LLOMe. Thapsigargin causes a sharp spike, whereas LLOMe produces minimal change, indicating the depletion of lysosomal calcium. Panel C shows galectin-3 staining in control and calcium-depleted cells treated with 0.25 mM LLOMe, quantified in panel D. Panel E shows normalized cytosolic SRG intensity upon 0.25 mM LLOMe with or without lysosomal calcium.
Ca 2+ efflux is not required for the protective mechanism conferred by TMEM63A. (A and B) GCaMP6s recordings from 20 to 30 WT RAW 264.7 cells. The change in fluorescence (ΔF) was normalized to the highest value for each cell. n = 3. (C–E) WT BMDM depleted of Ca2+ and challenged with LLOMe and quantified as in Fig. 1. n = 3; scale bars 10 µm.