Skip to main content
. Author manuscript; available in PMC: 2018 Oct 13.
Published in final edited form as: Circ Res. 2017 Aug 8;121(9):1081–1091. doi: 10.1161/CIRCRESAHA.117.311747

Figure 4. Expression of ADPR-insensitive TRPM2 mutant (C1008→A) in endothelial cells impairs Ca2+ entry and endotoxin-induced PMN uptake.

Figure 4

(A–B) HLMVECs were transfected with empty vector or ADPR-insensitive TRPM2 mutant (C1008→A) plasmid for 24h (A). Cells were then loaded with FLIPR-5 Ca2+ sensitive dye for 30 min, followed by stimulation with H2O2 (300 μM). H2O2-induced Ca2+ influx was suppressed in cells expressing the TRPM2 mutant (B). Representative tracing obtained with each experiment consisting of ≥6 replicates. (B) C57BL/6 mice were injected i.v. with liposomes containing empty vector or TRPM2 mutant plasmid for 48h followed by LPS challenge (10 mg/kg, i.p.) for 24h. Lungs were collected and subjected to immunoblot analysis of TRPM2 protein expression (B) and MPO activity (C). Expression of TRPM2 mutant suppressed the LPS-induced PMN uptake in lungs as determined by MPO activity (left panel) and H&E staining. n= 6 mice per group. Data are shown as mean ± SEM. *p<0.05 compared to the respective control group. Scale bar- 100μm.