Journal: bioRxiv
Article Title: Pharmacological CDK4/6 inhibition unravels a p53-induced secretory phenotype in senescent cells
doi: 10.1101/2020.06.05.135715
Figure Lengend Snippet: 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). N=6 mice/group. 14 dpt, bioluminescence was visualized and quantified by the IVIS spectrum in vivo imaging system, as shown by representative bioluminescence images ( A ) and quantification ( B ). ( C ) RNA isolated from treated kidneys and mRNA encoding p16 quantified by qRT-PCR. Representative images ( D ) to visualize SA-β-gal activities in vehicle-, doxorubicin-or abemaciclib-treated mouse kidney sections at 15 dpt (arrows indicated positive area; scale bar, 1 mm; N=3) and quantified ( E ). ( F ) 15 dpt, plasma was collected and expression levels of CXCL1 measured by ELISA. ( G ) Protein lysate obtained from drug treated kidneys to quantify IL6 by ELISA. ( H and I ) RNA isolated from kidneys and mRNA encoding indicated genes quantified by qRT-PCR. ( J ) Relative weight changes were calculated at 7 dpt and 14 dpt. Red blood cells ( K ) and white blood cells ( L ) were counted at 15 dpt. Percentage of T cells, B cells, granulocytes and macrophages were determined by flow cytometry analysis ( M ). Physical performance was measured by rotarods assay at 15 dpt ( N ), grip strength meter at 7 dpt and 14 dpt ( O ), and hanging tests were performed at 7 dpt and 14 dpt and normalized to weights ( P ). One-way ANOVA, data are means ±SD ( B, C, E, F, G, K, L and N ). Two-way ANOVA, data are means ±SD ( H, I, J, M, O and P ). *p<0.05, **p<0.01, ***p<0.001, N.S.=not significant. dpt, days post treatment.
Article Snippet: 20 μg total protein was collected from each treated kidney and IL6 protein level was determined by the mouse IL6 duo-set (R&D Systems) ELISA.
Techniques: In Vivo Imaging, Isolation, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, Flow Cytometry
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