Fig. 8: Inhibiting TRPM2 activation suppresses foam cell formation.

(a, b) Representative images and quantification of Oil Red O (ORO) staining of cultured macrophages after the treatment with oxLDL (50 μg/ml) for 24 h. 3 dishes of cells from 3 mice from each group were chosen for quantification. (c-f) Representative WB analysis and quantification of the expression of MCP1 and MIF in cultured macrophages treated with oxLDL (50 μg/ml) or TSP1 (10 μg/ml) for 24 h (n=3/group). (g, h) Inhibiting the activation of TRPM2 suppressed macrophage infiltration. in vitro macrophage infiltration test was performed as graphic illustration in Extended Data Fig. 3a (Red: F4/80; Blue: DAPI; Green: CD80). h, Quantification of the number of infiltrated macrophages within a x 10 field (n=6/group). (i, j) Inhibiting TRPM2 activation prevented the loss of emigration ability in oxLDL-pre-loaded macrophages. in vitro macrophage emigration test was performed as graphic illustration in Extended Data Fig. 3b. Macrophage emigration across endothelial cells induced by MCP1. Aorta-derived endothelial cells were plated on the transwell inserts (pore size: 12 μm) for 2–3 days. Macrophages preloaded with oxLDL for 24 h were added into the upper chamber after endothelial cells completely covered the upper surface of transwells. After 24 h, F4/80 and CD80 staining of macrophages in lower chamber was performed as in i (Red: F4/80; Blue: DAPI; Green: CD80). j, Quantification of the number of infiltrated macrophages within a x 10 field (n=6/group). (ns: no statistical significance; *: p < 0.05; **: p < 0.01; ***: p < 0.001; ANOVA, two-tailed, Bonferroni’s test; mean ± SEM).