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. 2020 Mar 19;295(19):6641–6651. doi: 10.1074/jbc.RA119.011192

Figure 2.

Figure 2.

TNFα or pervanadate treatment induces EMCN cleavage in HUVECs. a, schematic of the tagged EMCN and its cleavage. b, HUVECs were transfected with a construct encoding EMCN tagged with myc on the N terminus and DDK on the C terminus or with an empty construct as a control. Cell lysates were harvested 72 h after transfection for Western blot analysis. c and d, Western blotting confirmed the overexpression of tagged EMCN. HUVECs were transduced with AdEMCN or AdGFP as a control and serum-starved overnight. Cell lysates were harvested after treatment with TNFα (1 ng/ml, 24 h) or pervanadate (50 μm, 30 min) and processed for protein analysis. Western blot analysis for the DDK tag demonstrated the presence of EMCN-CTF in AdEMCN-transduced groups, which was absent in the AdGFP group. Treatment with TNFα or pervanadate led to an increase in EMCN-CTF. n = 3, Student's t test. *, p < 0.05; **, p < 0.01.