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. 2020 Aug 13;10:13708. doi: 10.1038/s41598-020-70126-7

Figure 2.

Figure 2

ATG7 knockdown inhibits thrombin-mediated expression of inflammatory proteins and NF-κB activity. HPAEC were transfected with si-Con or si-ATG7 for 48 h and then treated with thrombin (5 U/ml) for 6 h. (A,B) Conditioned media was collected from the cells and ELISAs were performed for (A) IL-6, (B) MCP-1. Error bars represent mean ± S.E. (n = 5 for each condition). (C&D) Cell lysates were analyzed by ELISA for (C) ICAM-1 and (D) VCAM-1 levels. Error bars represent mean ± S.E. (n = 5 for each condition). (E) HPAEC were transfected with si-Con or si-ATG7 for 24 h, then transfected with NF-κBLUC and Renilla LUC constructs as described in the “Materials and Methods”. The cells were then treated with thrombin (5 U/ml) for 6 h and cell extracts were assayed for Firefly and Renilla luciferase activities. Renilla luciferase was used as an internal control for transfection efficiency. Error bars represent mean ± S.E. (n = 6 for each condition).