Figure 1. TNFα-induces inhibition of cell proliferation and premature senescence in HUVECs.

(a) Experimental design: Upon propagation in full growth medium for 24 hours, cells were exposed to TNFα (10 ng/ml) ± inhibitors (NF-κB inhibitors plumericin (1.5 μM), PHA-408 (2 μM) and antioxidant NAC (2 mM)) for a period of six days, followed by a three days recovery period in full growth medium. Control HUVECs were grown in full growth medium only during the whole period. (b) Growth curves of HUVECs treated with or without TNFα for the indicated time points. (c) Increase in size and flattening of TNFα-treated cells. (d–f) Representative fluorescent images of HUVECs (day nine) grown in presence or absence of TNFα and their quantification: (d) Ki-67, (e) p21, and (f) p16.Values are presented as mean ± SD of technical triplicates (**p < 0.01, ***p < 0.001). The results shown are representative of three independent experiments.