Figure 3(A) 8 dpt, cells were stained for γ-H2AX (scale bar, 60 μm) and quantified (vehicle=288 cells, doxorubicin=280 cells, palbociclib=276 cells, abemaciclib=266 cells from 3 independent experiments). Data information: Data are means ±SD. One-way ANOVA *p<0.05, **p<0.01, ***p<0.001, N.S.= Not significant. Doxo, doxorubicin. Abe, abemaciclib. dpt, days post treatment. (B) BJ cells transduced with a NF-κB reporter were treated with vehicle, doxorubicin, paclitaxel (50 nM for 24 hours), palbociclib (1 μM for 8 times 24 hours) or abemaciclib and luciferase activity measured 8 dpt. TNFα treatment was used as positive control (n=3 independent experiments). Data information: Data are means ±SD. One-way ANOVA *p<0.05, **p<0.01, ***p<0.001, N.S.= Not significant. (C Venn plot of RNA-sequencing datasets of treated cells at 8 dpt (n=3 independent samples, sequenced together) (C). for doxorubicin- or abemaciclib-treated groups (8 dpt) relative to vehicle-treated group from the RNA-sequencing datasets (p-value<0.01 was regarded as significant). D) Heatmap of the SASP genes for doxorubicin- or abemaciclib-treated groups (8 dpt) relative to vehicle-treated group from the RNA-sequencing datasets (p-value<0.01 was regarded as significant). (E) 8 dpt, qRT-PCR of indicated NF-κB associated SASP genes was performed using indicated treated BJ cells 8 dpt (n=3 independent experiments). Data information: Data are means ±SD. Two-way ANOVA *p<0.05, **p<0.01, ***p<0.001, N.S.= Not significant. (F) Serum-free conditioned media (CM) was collected from vehicle-, doxorubicin-, paclitaxel-, palbociclib- or abemaciclib-treated BJ cells 8 dpt and expression levels of CXCL1 and CCL5 measured by ELISA (n=6 independent experiments). Data information: Data are means ±SD. One-way ANOVA *p<0.05, **p<0.01, ***p<0.001, N.S.= Not significant. (G) Proteins in the serum-free conditioned media (CM) collected from doxorubicin- or abemaciclib-treated BJ fibroblasts (8 dpt) were measured by cytokine array. Purple boxes indicated internal positive controls from each membrane; Red boxes highlighted the most typical SASP factors. p16-3MR mice were treated with vehicle (PBS, 7 consecutive days), doxorubicin (5 mg/kg, 3 consecutive days) or abemaciclib (50 mg/kg, 7 consecutive days). 15 dpt, plasma was collected and expression levels of CXCL1 measured by ELISA (H) Data information: Data are means ±SD. One-way ANOVA *p<0.05, **p<0.01, ***p<0.001, N.S.= Not significant. p16-3MR mice were treated with vehicle (PBS, 7 consecutive days), doxorubicin (5 mg/kg, 3 consecutive days) or abemaciclib (50 mg/kg, 7 consecutive days). protein lysate obtained from drug treated kidneys to quantify IL6 by ELISA (I), Data information: Data are means ±SD. One-way ANOVA *p<0.05, **p<0.01, ***p<0.001, N.S.= Not significant. p16-3MR mice were treated with vehicle (PBS, 7 consecutive days), doxorubicin (5 mg/kg, 3 consecutive days) or abemaciclib (50 mg/kg, 7 consecutive days). RNA isolated from kidneys and mRNA encoding indicated NF-κB associated genes (J) quantified by qRT-PCR. n=6 mice/group. Data information: Data are means ±SD. Two-way ANOVA *p<0.05, **p<0.01, ***p<0.001, N.S.= Not significant. p16-3MR mice were treated with vehicle (PBS, 7 consecutive days), doxorubicin (5 mg/kg, 3 consecutive days) or abemaciclib (50 mg/kg, 7 consecutive days). From a subset of the mice, RFP+ cells were sorted from the renal cortex using FACS and mRNA encoding indicated NF-κB associated pro-inflammatory genes quantified by qRT-PCR. The values of fold change over vehicle were plotted in the heatmap. n=4 mice/group (K). (L) Cell-free plasma was derived from breast cancer patients treated with paclitaxel (n=10) or palbociclib (n=9) and the expression levels of CXCL1, CCL2, CCL5 and MMP1 quantified by ELISA. The values represent the ratio between on-treatment and pre-treatment levels for each patient. Data information: Data are means ±SD. Unpaired student t-test (L). *p<0.05, **p<0.01, ***p<0.001, N.S.= Not significant.