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. Author manuscript; available in PMC: 2022 Sep 28.
Published in final edited form as: Nat Cardiovasc Res. 2022 Mar 28;1(4):344–360. doi: 10.1038/s44161-022-00027-7

Fig. 4: Deletion of Trpm2 inhibits foam cell formation.

Fig. 4:

(a, b) Representative images and quantification of ORO staining in macrophages from wild-type (WT) or Trpm2 knockout (M2KO) mice after 24h oxLDL treatment (n=6/group). (c, d) Representative WB analysis (WB) of CD36, pFyn, pJNK and pp38 expression in macrophages (n=6/group). (e, f) Representative images and quantification of ORO staining of macrophages (n=8/group) treated with and without anisomycin (10 μM), a p38 and pJNK activator. (g-h) Representative WB of CD36, pFyn, pJNK and pp38 expression in macrophages. BAPTA-AM (1 μM), an intracellular Ca2+ chelator. (i, j) Representative images and quantification of ORO staining of macrophages treated with BAPTA-AM (n=3/group). (k, l) k, Representative images of Rhodamine-123 (R123) real-time imaging before and 5 min after oxLDL treatment in macrophages. Control group (PBS treatment) was used to show the rapid photo bleaching of R123. l, Quantification of R123 fluorescence changes 5 min after oxLDL treatment (n=30,40,40,43, respectively). (m, n) Quantification and representative WB of iNOS expression in macrophages (n=6/group). (o, p) o, Representative real-time ratio Ca2+ imaging traces during oxLDL treatment. The averaged traces were from 10 macrophages randomly chosen from a representative culture dish for each group. p, Quantification of Fura-2 fluorescence changes 5 min after oxLDL treatment (n=20/group). (q, r) q, Representative images of R123 imaging before and 5 min after oxLDL treatment in macrophages with DMSO or BAPTA-AM (1 μM) preloading for 30 min. r, Quantification of R123 fluorescence changes 5 min after oxLDL treatment (n=20/group). (s) Measurement of IL-1β level in culture medium of isolated macrophages after oxLDL treatment using ELISA (n=4/group). (t, v) Representative WB of MCP1 and MIF expression in macrophages (n=6/group). (u, w) Representative WB of pp65 expression in macrophages (n=6/group). (oxLDL (50 μg/ml) was treated for 24 h for all the ORO staining and WB analysis; ns: no statistical significance; *: p < 0.05; **: p < 0.01; ***: p < 0.001; ANOVA, two-tailed, Bonferroni’s test; mean ± SEM).