Extended Data Fig. 2: Validation of cell-selective Trpm7 null mice.
From: The TRPM7 chanzyme in smooth muscle cells drives abdominal aortic aneurysm in mice
(a) Mouse tail genotyping of SMC-selective Trpm7 KO mice. n = 6. (b) PCR of aortic medial genomic DNA with primers flanking exon 17. The Trpm7sm-WT mice were treated with the vehicle (corn oil) or tamoxifen (Tam.). n = 3. (c) RT-qPCR of aortic media-derived mRNA with primers flanking exon 17. Residual band in the KO mice denotes undecayed truncated TRPM7 mutant transcript. n = 3. (d) Immunostaining of TRPM7 protein in abdominal aorta. Colocalization of TRPM7 with SMC as labeled by anti-SM22α was observed in wild type but absent in KO mice. Scale bar = 100μm. n = 3. (e-f) Immunoblots of TRPM7 protein in abdominal aorta from wild type or KO mice. Both full length TRPM7 (TRPM7-FL, ~220 kDa) or cleaved kinase (M7CK, ~65 kDa) were detected in wild type mice but abolished in KO mice. TRPM7-FL and M7CK were immunoblotted separately with anti-TRPM7 antibody commercially from Cayman Chemical and Sigma, respectively. n = 3. (g-h) Whole-cell patch clamping of TRPM7 currents in isolated aortic smooth muscle cells. Current density collected at 100 mV and the representative current traces are shown. n = 8 ~ 10. (i) Mouse tail genotyping of macrophage-selective Trpm7 KO mice. n = 6. (j) Immunofluorescent detection of TRPM7 protein in peritoneal macrophages. Scale bar = 200μm. n = 3. (k) Mouse tail genotyping of endothelial cell-selective KO mice. n = 6. (l) Immunofluorescent staining of TRPM7 in aortic endothelium. Colocalization of TRPM7 with endothelial marker (CD31) was seen in wild type mice but undetectable in KO mice. Scale bar = 100μm. n = 3. Unpaired 2-tailed Student’s t test was performed to compare two groups (Extended Data Figs. 2f, h).
