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. Author manuscript; available in PMC: 2021 Aug 1.
Published in final edited form as: Shock. 2020 Aug;54(2):245–255. doi: 10.1097/SHK.0000000000001445

Figure 2. MKT-077 protects against thrombin and LPS-induced inflammatory responses.

Figure 2.

Figure 2.

(A–D) HPAEC were treated with 1µg/ml MKT-077(1 h) followed by thrombin treatment (5 U/ml) for 6 h. Total cell lysate was immunoblotted for ICAM-1, VCAM-1, MCP-1 and IL-6 and culture supernatants were analyzed for IL-6 and MCP-1 using ELISA. (E) HPAEC were transfected with NF-κB-LUC and pTKRLUC constructs by DEAE-dextran as described in Methods. Cells were then treated with 1µg/ml MKT-077(1 h) before or 1 h after thrombin treatment (5 U/ml) for 6 h and cell extracts were prepared and assayed for Firefly and Renilla luciferase activities. (F&G) luciferase lysates were immunoblotted for IL-6 using ELISA and for ICAM-1 using anti-ICAM-1 antibody. Anti-GAPDH antibody was used to monitor loading control. (H–I) HPAEC were treated with 1µg/ml MKT-077 (1 h) followed by LPS treatment (0.1µg/ml) for 6 h. Cell culture supernatants were analyzed for IL-6 using ELISA. Total cell lysates were immunoblotted for ICAM-1 and VCAM-1. Anti-GAPDH antibody was used to monitor loading control. The blot in 2I is cropped from a gel.