Journal: bioRxiv

Article Title: A feed-forward loop between niche adenosine and Gzmk⁺ CD8 T cells propagates systemic inflammaging

doi: 10.64898/2026.03.18.712515

Figure Lengend Snippet: (A-D) The analysis of male 19 months WT or Gzmk -/- mice (male). (A) Staining of SA-β-gal in liver and hippocampus. (B) H&E staining in liver, lung and kidney. (C) SA-β-gal staining for WAT from indicated mice. (D) Western blot analysis of the expression of p16, p21 and p53 in liver. (E) The bioluminescence imaging of indicated mice (15 months, female) by injecting luciferase substrates. (F-G) CD8 T cells (4 x10 6 ) from old or old Gzmk -/- mice were adoptively transferred into young p16 Ink4a -luciferase reporter mice, one month later, the luciferase activity was measured (F), and the expression of the p16 -driven luciferase reporter in indicated tissues were measured by RT-qPCR.

Article Snippet: Primary antibodies used in this study: Mouse anti-CDKN2A/p16 (Santa Cruz Biotechnology, cat# sc-1661, 1:1000) Mouse anti-CDKN1A p21 (Santa Cruz Biotechnology, cat# sc-6246, 1:1000) Mouse anti-p53 (Santa Cruz Biotechnology, cat# sc-98, 1:1000) Mouse anti-GAPDH (Absin, abs830030,1:10000) Rabbit anti-γH2A.X (Cell Signaling Technology, cat# 9718S, 1:1000)

Techniques: Staining, Western Blot, Expressing, Imaging, Luciferase, Activity Assay, Quantitative RT-PCR

Journal: bioRxiv

Article Title: A feed-forward loop between niche adenosine and Gzmk⁺ CD8 T cells propagates systemic inflammaging

doi: 10.64898/2026.03.18.712515

Figure Lengend Snippet: Old mice (19 months) were administrated with SCH or PPACK for one-month. The level of IL-6 and TNFα (A), ALT and AST (B) in plasma from indicated mice were determined by ELSIA. (C) Staining of SA-β-gal in liver. (D) H&E staining of lungs from indicated mice. (E) Staining of SA-β-gal in hippocampus from indicated mice. (F-G) Western blot analysis of P16, P21 and P53 at liver and brain from indicated mice.

Article Snippet: Primary antibodies used in this study: Mouse anti-CDKN2A/p16 (Santa Cruz Biotechnology, cat# sc-1661, 1:1000) Mouse anti-CDKN1A p21 (Santa Cruz Biotechnology, cat# sc-6246, 1:1000) Mouse anti-p53 (Santa Cruz Biotechnology, cat# sc-98, 1:1000) Mouse anti-GAPDH (Absin, abs830030,1:10000) Rabbit anti-γH2A.X (Cell Signaling Technology, cat# 9718S, 1:1000)

Techniques: Clinical Proteomics, Staining, Western Blot

Journal: bioRxiv

Article Title: A feed-forward loop between niche adenosine and Gzmk⁺ CD8 T cells propagates systemic inflammaging

doi: 10.64898/2026.03.18.712515

Figure Lengend Snippet: (A-D) The analysis of male 19 months WT or Gzmk -/- mice (male). (A) Staining of SA-β-gal in liver and hippocampus. (B) H&E staining in liver, lung and kidney. (C) SA-β-gal staining for WAT from indicated mice. (D) Western blot analysis of the expression of p16, p21 and p53 in liver. (E) The bioluminescence imaging of indicated mice (15 months, female) by injecting luciferase substrates. (F-G) CD8 T cells (4 x10 6 ) from old or old Gzmk -/- mice were adoptively transferred into young p16 Ink4a -luciferase reporter mice, one month later, the luciferase activity was measured (F), and the expression of the p16 -driven luciferase reporter in indicated tissues were measured by RT-qPCR.

Article Snippet: Primary antibodies used in this study: Mouse anti-CDKN2A/p16 (Santa Cruz Biotechnology, cat# sc-1661, 1:1000) Mouse anti-CDKN1A p21 (Santa Cruz Biotechnology, cat# sc-6246, 1:1000) Mouse anti-p53 (Santa Cruz Biotechnology, cat# sc-98, 1:1000) Mouse anti-GAPDH (Absin, abs830030,1:10000) Rabbit anti-γH2A.X (Cell Signaling Technology, cat# 9718S, 1:1000)

Techniques: Staining, Western Blot, Expressing, Imaging, Luciferase, Activity Assay, Quantitative RT-PCR

Journal: bioRxiv

Article Title: A feed-forward loop between niche adenosine and Gzmk⁺ CD8 T cells propagates systemic inflammaging

doi: 10.64898/2026.03.18.712515

Figure Lengend Snippet: (A, B) Primary human bladder fibroblasts were isolated from surgical excision from bladder cancer patients and subjected to GZMK treatment. (A) HBFs were treated with 100nM GZMK for 5 days, and SA-β-gal staining were performed. (B) HBFs were treated with 100nM GZMK in medium without FBS for one day, IL-6 and TNFα in supernatant were measured by ELISA. (C, D) MEFs were isolated and subjected to GZMK treatment as same with . (E-H) BMDMs were subjected to Gzmk or indicated inhibitors treatment. (E) BMDMs were stimulated with Gzmk for 30 minutes and phosphorylated ERK and p38 were determined by flow cytometry. (F) BMDMs were cultured with Gzmk or combined with indicated inhibitors for three days, SA-β-gal staining were performed. BMDMs were cultured with Gzmk or combined with indicated inhibitors for one day, expression of P16 (G) and P21 (H) were analyzed by flow cytometry, IL-6 and TNFα (I) in supernatant were measured by ELISA.

Article Snippet: Primary antibodies used in this study: Mouse anti-CDKN2A/p16 (Santa Cruz Biotechnology, cat# sc-1661, 1:1000) Mouse anti-CDKN1A p21 (Santa Cruz Biotechnology, cat# sc-6246, 1:1000) Mouse anti-p53 (Santa Cruz Biotechnology, cat# sc-98, 1:1000) Mouse anti-GAPDH (Absin, abs830030,1:10000) Rabbit anti-γH2A.X (Cell Signaling Technology, cat# 9718S, 1:1000)

Techniques: Isolation, Staining, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Cell Culture, Expressing

Journal: bioRxiv

Article Title: A feed-forward loop between niche adenosine and Gzmk⁺ CD8 T cells propagates systemic inflammaging

doi: 10.64898/2026.03.18.712515

Figure Lengend Snippet: Old mice (19 months) were administrated with SCH or PPACK for one-month. The level of IL-6 and TNFα (A), ALT and AST (B) in plasma from indicated mice were determined by ELSIA. (C) Staining of SA-β-gal in liver. (D) H&E staining of lungs from indicated mice. (E) Staining of SA-β-gal in hippocampus from indicated mice. (F-G) Western blot analysis of P16, P21 and P53 at liver and brain from indicated mice.

Article Snippet: Primary antibodies used in this study: Mouse anti-CDKN2A/p16 (Santa Cruz Biotechnology, cat# sc-1661, 1:1000) Mouse anti-CDKN1A p21 (Santa Cruz Biotechnology, cat# sc-6246, 1:1000) Mouse anti-p53 (Santa Cruz Biotechnology, cat# sc-98, 1:1000) Mouse anti-GAPDH (Absin, abs830030,1:10000) Rabbit anti-γH2A.X (Cell Signaling Technology, cat# 9718S, 1:1000)

Techniques: Clinical Proteomics, Staining, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: Targeting EphA2 under DNA damage causes mitotic bypass via p21 induction

doi: 10.1016/j.jbc.2026.111271

Figure Lengend Snippet: p21 is responsible for EphA2 knockdown–caused mitotic bypass. A and E , HeLa S3 cells were transfected with either siCtrl or siEphA2. After 12 h, the cells were treated with 0.5 μM ADR for 12 h, washed, and then further cultured for 48 h before Western blot analysis ( A ). Band intensities of cyclin B1 ( B ), p53 ( C ), and p21 ( D ) were quantified, and the ratios against the band intensity of α-tubulin are plotted. Three or four independent experimental results are shown as dots together with mean ± SD. p -values were determined using Welch's t test. E , HeLa S3 cells were transfected with either nontargeting or p21-targeting siRNA and further cultured for 48 h before Western blot analysis. At 24 h after transfection, the medium was changed with fresh medium. F and G , HeLa S3/tFucci(CA)5 cells were transfected with siEphA2, along with either siCtrl or sip21. After 12 h, the cells were treated with 0.5 μM ADR for 12 h, washed, and then further cultured for 24 h before time-lapse imaging for 24 h. G2 cells at the beginning of imaging were examined. In F , percentages of cells in G2 phase, cell death during G2 phase, and the cumulative percentage of mitotic bypass are shown as mean ± SD from three independent experiments. No cells entered M phase. G2, orange ; M/post-M, gray ; cell death, black ; mitotic bypass, blue . Percentages of mitotic bypass during imaging are shown in the graph ( G ) as mean ± SD calculated from three independent experimental results. p -values were determined using Student's t test. n = 40 cells per condition. ADR, adriamycin.

Article Snippet: Primary antibodies used for immunoblotting (IB), immunofluorescence (IF) or flow cytometry were as follows: mouse monoclonal anti-Chk1 (IB, 1:1000; G-4, sc-8408, Santa Cruz Biotechnology), anti-γ-tubulin (IF, 1:250; GTU-88, T6557, Merck), anti-p21 (IB, 1:1000, DCS60, 2946, Cell Signaling Technology), anti-p53 (IB, 1:1000; DO-1, sc-126, Santa Cruz Biotechnology), and anti-phospho-histone H3 (IF, 1:400; 6G3, 9706, Cell Signaling Technology) antibodies; rabbit monoclonal anti-EphA2 (IB, 1:1000; 6997S, Cell Signaling Technology), anti-phospho-Chk1 (Ser345, IB, 1:1000; 133D3, #2348, Cell Signaling Technology), and anti-phospho-EphA2 (Ser897, IB, 1:1000; D9A1, 6347, Cell Signaling Technology) antibodies; rabbit polyclonal anti-cyclin B1 (IB, 1:3000; IF and flow cytometry, 1:250; H-433, sc-752, Santa Cruz Biotechnology), anti-phospho-histone H2A.X (γH2AX, IB, 1:500; 2577S, Cell Signaling Technology), and anti-phospho KAP1 (Ser824, IB, 1:1000; A300–767A, Bethyl Laboratories, Montgomery) antibodies; and rat monoclonal anti-α-tubulin (IB, 1:4000; IF, 1:800; MCA78 G, Bio-Rad) antibody.

Techniques: Knockdown, Transfection, Cell Culture, Western Blot, Imaging

Journal: The Journal of Biological Chemistry

Article Title: Targeting EphA2 under DNA damage causes mitotic bypass via p21 induction

doi: 10.1016/j.jbc.2026.111271

Figure Lengend Snippet: p21 upregulation induced by EphA2 knockdown in human cervical cancer cells. Cells were transfected with either siCtrl or siEphA2. After 12 h, cells were treated with ADR for 12 h, washed with PBS (−), and further cultured for 48 h prior to Western blot analysis. Ca Ski cells ( A ), SKG-II cells ( C ), HCT116 cells ( D ), and MCF7 cells ( E ) were subjected to this using the ADR concentrations indicated in the figure. Ca Ski cells were additionally subjected to time-lapse imaging during the last 24 h of the 48 h culture period. The same procedure was applied to Ca Ski cells, and time-lapse imaging was performed during the last 24 h of a 48 h culture ( B ). Representative results were shown from two independent results. Percentages of cells in G2 phase, M/post-M, cell death in G2 phase and the cumulative percentage of mitotic bypass at the time points are shown. n = 40 cells per condition. ADR, adriamycin.

Article Snippet: Primary antibodies used for immunoblotting (IB), immunofluorescence (IF) or flow cytometry were as follows: mouse monoclonal anti-Chk1 (IB, 1:1000; G-4, sc-8408, Santa Cruz Biotechnology), anti-γ-tubulin (IF, 1:250; GTU-88, T6557, Merck), anti-p21 (IB, 1:1000, DCS60, 2946, Cell Signaling Technology), anti-p53 (IB, 1:1000; DO-1, sc-126, Santa Cruz Biotechnology), and anti-phospho-histone H3 (IF, 1:400; 6G3, 9706, Cell Signaling Technology) antibodies; rabbit monoclonal anti-EphA2 (IB, 1:1000; 6997S, Cell Signaling Technology), anti-phospho-Chk1 (Ser345, IB, 1:1000; 133D3, #2348, Cell Signaling Technology), and anti-phospho-EphA2 (Ser897, IB, 1:1000; D9A1, 6347, Cell Signaling Technology) antibodies; rabbit polyclonal anti-cyclin B1 (IB, 1:3000; IF and flow cytometry, 1:250; H-433, sc-752, Santa Cruz Biotechnology), anti-phospho-histone H2A.X (γH2AX, IB, 1:500; 2577S, Cell Signaling Technology), and anti-phospho KAP1 (Ser824, IB, 1:1000; A300–767A, Bethyl Laboratories, Montgomery) antibodies; and rat monoclonal anti-α-tubulin (IB, 1:4000; IF, 1:800; MCA78 G, Bio-Rad) antibody.

Techniques: Knockdown, Transfection, Cell Culture, Western Blot, Imaging

Journal: Nature Communications

Article Title: Engineered exosome nanovesicles for delivery of antibodies to treat inflammatory bowel disease

doi: 10.1038/s41467-026-69382-4

Figure Lengend Snippet: a , b During the treatment of AOM/DSS-induced CAC mice with PrEXO-a23 or PrEXO-a23 and the p53 inhibitor (ip53), body weight ( a ) and DAI score ( b ) of mice were monitored daily. c spleen index analysis of mice. d – g Colon photographs ( d ), colon length ( e ), tumor number ( f ), tumor burden ( g ) of CAC mice at week 10. h Relative mRNA expression levels of Trp53 , Cdkn1a and Bax in colonic tissues of CAC mice with indicated treatment measured by RT-qPCR assay. i , j Western blot images ( i ) and relative protein level analysis ( j ) of p53, p21, and Bax expressed in colonic tissues. For ( a – c , e – h , j ), data were presented as mean ± SD ( n = 4 mice in a – c , e – g ; n = 5 mice in h ; n = 3 mice in j ). For ( c , e – h , j ), statistical significance was determined using one-way ANOVA with Tukey’s multiple comparisons test. Source data are provided as a Source Data file.

Article Snippet: Anti-mouse p21 and Bax were obtained from Cell Signaling Technology.

Techniques: Expressing, Quantitative RT-PCR, Western Blot