Fig. 1.

Accumulation set-up assay. Micro-vessels of HUVECs cultured against ECM, were treated with different concentrations of TBHP (0, 0.01, 0.1, 1, 3, 5, 10 and 20 mM) for 0.5 h and stained for DNA with Hoechst (blue), for ROS with DHR123 (green) and dead cells with PI (red) (A) Representative images of bottom part of the micro-vessel of HUVEC (i) sum-projection of all condition (ii) Max-projection of a part of micro-vessel treated with TBHP 10 mM captured at 20x magnification. Scale bar is 100 μm (i) and 40 μm (ii). Graphs of (B) Cells count (C) cell viability expressed as percentage (D) ROS content in a log scale expressed as fluorescence intensity normalized against the number of cells count (E) Non-linear regression analysis on ROS level over log scale concentration of TBHP. The table above the graph includes information of the model to fit the data. Data are expressed as average ± standard deviation (STD) with N = 3 and n = 2–10 replicates. Difference between results were evaluated by one-way ANOVA with Tukey's multiple comparison (graph in B) or Dunnett's test (graph in D). P-value < 0.05 was considered statistically significant * p < 0.05, ****p < 0.0001. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)