fig2

Figure 2. Senolytics D+Q rescue iPSC-CM number and cell cycle activity. (A and B) Percent of crystal violet stained iPSC-CMs (A) and BrdU-positive iPSC-CMs (B) when cultured alone (CTRL 17d), co-cultured with senCPCs (Co-culture 17d) or co-cultured with senCPCs and treated with D+Q (Co-culture D+Q 17d). Data are Mean ± SD. Significance was determined by ANOVA followed by Tukey’s post-hoc test to identify the differences. *P < 0.05, **P < 0.01, ****P < 0.0001. Individual data points represent independent replicates/wells. Representative field of view micrograph images of iPSC-CMs stained with crystal violet (A). Scale bar = 200 μm. Representative field of view micrograph images of iPSC-CMs labelled and immunostained with BrdU (green) and nuclei counterstained with DAPI (blue) (B). Scale bar = 50 μm. (C and D) SA-β-gal activity (B) and p21 expression (D) of iPSC-CMs when cultured alone (CTRL 17d), co-cultured with senCPCs (Co-culture 17d) or co-cultured with senCPCs and treated with D+Q (Co-culture D+Q 17d). Data are Mean ± SD. Individual data points represent independent replicates/wells. Representative field of view micrograph images of iPSC-CMs stained by enzymatic SA-β-gal assay (blue, upper panel) and nuclei counterstained with DAPI (blue, lower panel) (C). Scale bar = 100 μm. Representative field of view micrograph images of iPSC-CMs immunostained for p21 (red) and nuclei counterstained with DAPI (blue) (D). Scale bar = 50 μm.