Figure 7.

Prolonged interaction of UCB with the HBMEC monolayer activates autophagy. HBMEC line was incubated without (control) or with 50 or 100 μM UCB, in the presence of 100 μM HSA, for the indicated periods. Autophagy was evaluated by Western blot, with LC3 I (free form) and II (phosphatidylethanolamine-conjugated form) detection. Representative results of one experiment are shown (A). Quantification of LC3 I and II band intensity was done by scanning densitometry and the ratio between the membrane-bound and the cytosolic forms was calculated and standardized with respect to β-actin protein (B). Results are means ± SEM from at least four independent experiments performed in duplicate. ^*P < 0.05 and ^**P < 0.01 vs. control. ^##P < 0.01 UCB 100 vs. 50 μM.