N-Octanoyl Dopamine Treatment of Endothelial Cells Induces the Unfolded Protein Response and Results in Hypometabolism and Tolerance to Hypothermia
Figure 5
NOD does not impair cell viability.
(a) HUVECs were treated for 24 hrs with NOD (100 µM) or tunicamycin (TM, 1 µg/ml). In addition cells were treated for 3 hrs with 100 µM of NOD prior to tunicamycin treatment. Cells that were not treated (Med.) served as control. Both floating and adherent cells were harvested, stained for Annexin V and 7-AAD and analysed by flow cytometry. (b) HUVECs were treated with 100 µM NOD or TM (1 µg/ml) for the indicated time periods. Total RNA was isolated and expression of CHOP mRNA was assessed by qPCR and normalized for GAPDH (left panel). Cells were pre-treated either with NOD or with tunicamycin and then chased for 0, 2, 4 and 8 hrs with AcD 50 ng/ml. Total RNA was isolated and the expression of CHOP (middle panel) and BiP (right panel) mRNA were assessed by qPCR and normalized for GAPDH. The results of three different experiments are expressed as mean mRNA levels ± SD, ** P<0.01 vs. untreated cells.