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Figure 3. Senolytics D+Q conditioned media rescue HUVEC number, migration and tube formation. (A and B) Percent of crystal violet stained HUVECs (A) and Ki67-positive HUVECs (B) when cultured in normal growth media (GM), senCPC conditioned media (SenCPC CM), and D+Q conditioned media (D+Q CM). Data are Mean ± SD. Significance was determined by ANOVA followed by Tukey’s post-hoc test to identify the differences. ****P < 0.0001, *P < 0.05. Individual data points represent independent replicates/wells. Representative field of view micrograph images of HUVECs stained with crystal violet (A) and Ki67 (red; B). Nuclei counterstained with DAPI (blue). Scale bar = 50 μm. (C) HUVEC migration measured by the scratch assay when supplemented with normal growth media (GM), senCPC conditioned media (SenCPC CM), and D+Q conditioned media (D+Q CM) for 24 h. Data are Mean ± SD% of control (GM). Significance was determined by ANOVA followed by Tukey’s post-hoc test to identify the differences ****P < 0.0001, **P < 0.01. Representative field of view micrograph images of the scratch (red dotted lines) in the different media conditions. (D) Total tube length formed by HUVECs when cultured in normal growth media (GM), senCPC conditioned media (Sen-CPC CM), and D+Q conditioned media (D+Q CM). Data are Mean ± SD% of control (GM). Significance was determined by ANOVA followed by Tukey’s post-hoc test to identify the differences. *P < 0.05. Individual data points represent independent replicates/wells. Representative field of view micrograph images of HUVEC tube formation. Scale bar = 200 μm.