Figure S1 - HLA-B35 and dsRNA Induce Endothelin-1 via Activation of ATF4 in Human Microvascular Endothelial Cells

Treatment with mouse monoclonal anti-MHC class I blocking antibody did not affect ET-1 and ATF4 mRNA levels in HDMECs transfected with HLA-B35. PPET-1 (right panel) and ATF4 (left panel) mRNA after HLA-B35 (or HLA-B8) treatments with increasing dose of mouse monoclonal anti-MHC class I blocking antibody (W6/32) in HDMECs. Confluent dishes of HDMECs were transduced with 10 MOI of Adenovirus encoding HLA-B35/GFP (Ad-HLA-B8/GFP) for 24 h and then with increasing dose of W6/32 (0.1–10 µg/ml) for the following 24 hours. Total RNA was extracted and mRNA levels of PPET-1 and ATF4 were quantified by quantitative RT-PCR. Expression of the housekeeping gene β-actin served as an internal positive control in each assay performed. After measurement of the relative fluorescence intensity for each sample, the amount of each mRNA transcript was expressed as a threshold cycle value. (TIF)