Fig. 5. Protein expression of eNOS_Ser1177 and total eNOS in endothelial cells treated with TNFα and EMPA after 6 hours. Cells were treated with 0.02% DMSO (control), 10 ng/mL TNFα or 10 ng/mL TNFα with 1 µM EMPA. eNOS_Ser1177 and total eNOS levels were determined after 6h TNFα stimulation in HCAECs (a+b, g, n=6) and HUVECs (d+e, g n=6). eNOS_Ser1177 phosphorylation was determined by the ratio of eNOS_Ser1177/eNOS (c,f). Representative images of Western blots for eNOS_Ser1177 (g) together with total eNOS and GAPDH. Unrelated bands omitted for clarity. All full length blots can be found in the supplemental material. Data are presented as mean±SD. *p<0.05 vs. TNFα