Skip to main content
. 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 3. Mortalin knockdown attenuates thrombin-induced NF-κB reporter activity and mitigates inflammatory gene expression.

Figure 3.

(A) HPAEC grown to confluence on 2% gelatin-coated coverslips were transfected with control siRNA or mortalin siRNA using DharmaFect1. After 24–36 h cells were fixed, permeabilized and stained with anti-mortalin antibody to visualize depletion of mortalin. (B) Total cell lysates were immunoblotted for mortalin levels using anti-mortalin antibody. (C-F) HPAEC were transfected with control siRNA or mortalin siRNA using DharmaFect1. Twenty four hours later cells were again transfected with NF-κB-LUC and pTKRLUC constructs by DEAE-dextran as described in Methods. Cells were then challenged with thrombin (5 U/ml) for 6 h. Cell extracts were prepared and assayed for Firefly and Renilla luciferase activities. Lysates were also immunoblotted for ICAM-1 and VCAM-1 using anti-ICAM-1 and anti-VCAM-1 antibody. Anti-RelA/p65 antibody was used as a loading control.