FIGURE 2.

Metformin decreased DOX-induced SASP factors in conditioned media of HUVECs. HUVECs were treated for 24 h with 0.5 µM DOX ± metformin (2 and 5 mM, added 24 h before DOX). Thereafter, DOX was removed and the cells were incubated in DOX-free media with or without metformin for an additional 72 h. Conditioned media were collected and the protein expression of SASP factors including IL-6, TNF-α, MIP-1α, MCP-3, MCP-1, CXCL1, CXCL2, IL-1 β, and IL-8 [(A–I), respectively (n = 5–8)] was determined by Luminex multiplex platform. Values were normalized to the protein concentration of the cells determined by BCA. Values are shown as means ± SEM. Data were analyzed by one-way ANOVA followed by a Dunnet’s multiple comparisons test (Figures 2A,B, D–E, G, I) or non-parametric Kruskal–Wallis tests followed by Dunn’s post hoc test (Figures 2C,F,H); * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.