dy406 05 duo set  (R&D Systems)

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

Journal: Frontiers in Molecular Neuroscience

Article Title: Transcriptional Profiling of Non-injured Nociceptors After Spinal Cord Injury Reveals Diverse Molecular Changes

doi: 10.3389/fnmol.2019.00284

Figure Lengend Snippet: Behavioral responses and cytokines. (A1) Open field behavior (10 min trials) in naïve or sham mice 0, 1, 3, 5 and 7 days post-surgery does not differ significantly at any time point in total ambulation, N = 3–10 sham, 4 naïve. (A2) Total ambulation differs significantly 1 day post-spinal cord injury (SCI) in both SCI vs. naïve (one-way ANOVA, p = 0.0059, Tukey’s multiple comparisons test, ** p < 0.005) and SCI vs. sham (* p < 0.05) mice. (B1) Mechanical and (B2) thermal sensitivity do not differ significantly 0, 1, 3, 5, and 7 days post-surgery in naïve and sham mice, N = 6 each. (C) Cytokine ELISAs on spinal cord segments at the level of laminectomy (T8–T11) show no significant differences between naïve and sham mice 1 or 4 days post-surgery. One-day post-operation, SCI mice have significantly increased levels of IL-6 and IL-10 compared to naïve controls (one-way ANOVA, Bonferroni’s multiple comparisons test; * p < 0.05, ** p < 0.005, respectively) and significantly increased levels of IL-10 in comparison to 4 days sham condition (one-way ANOVA, Bonferroni’s multiple comparisons test; ∧∧∧ p < 0.001). Cuprizone-treated mice as positive control, with significantly increased TNF-α, IL-1β, and IL-10 compared to naïve controls (one-way ANOVA, Bonferroni’s multiple comparisons test; * p < 0.05, ** p < 0.005, ∧∧∧ p < 0.001, respectively) after 5 weeks of cuprizone treatment and significant increases in IL-1β at the 1 day sham time point, and IL-10 at the 4 days sham time point (one-way ANOVA, Bonferroni’s multiple comparisons test; * p < 0.05, ∧∧∧∧ p < 0.0001, respectively). *Represents comparisons to naïve conditions, # represents comparisons to 1 day sham conditions, ∧ represents comparisons to 4 days sham conditions.

Article Snippet: The ELISA assays were performed according to the manufacturer’s instructions (R&D systems mouse duo-sets IL-10, IL-6, IL-1β, TNF-α, completed with Ancillary Reagent Kit 2, Minneapolis, MN, USA).

Techniques: Comparison, Positive Control