Journal: Cancer Reports

Article Title: ANXA1 / FPR Signaling Contribute to Paclitaxel Resistance in Ovarian Cancer Through PI3K / AKT ‐Associated PGC1α Regulation

doi: 10.1002/cnr2.70637

Figure Lengend Snippet: Antibodies.

Article Snippet: Annexin A1 , ABclonal , A1118 , 1:1000.

Techniques:

Journal: Cancer Reports

Article Title: ANXA1 / FPR Signaling Contribute to Paclitaxel Resistance in Ovarian Cancer Through PI3K / AKT ‐Associated PGC1α Regulation

doi: 10.1002/cnr2.70637

Figure Lengend Snippet: Sequence of related primers is as follows.

Article Snippet: Annexin A1 , ABclonal , A1118 , 1:1000.

Techniques: Sequencing

Journal: Cancer Reports

Article Title: ANXA1 / FPR Signaling Contribute to Paclitaxel Resistance in Ovarian Cancer Through PI3K / AKT ‐Associated PGC1α Regulation

doi: 10.1002/cnr2.70637

Figure Lengend Snippet: High expression of ANXA1 may be related to paclitaxel sensitivity of ovarian cancer cells. (A) The relationship between the expression of ANXA1 and the survival of ovarian cancer patients was analyzed by the TCGA database. (B) The expression of ANXA1 was analyzed by immunohistochemistry results of ovarian cancer clinical samples in The Human Protein Atlas database (40×). (C) Dose–response curves of A2780 and A2780/PA cells treated with different concentrations of paclitaxel. IC50: 0.664 μg/mL (A2780) and 31.47 μg/mL (A2780/PA). (D) ANXA1 mRNA expression of A2780 and A2780/PA was detected by qPCR assay. (E) The protein expression of ANXA1 in A2780 and A2780/PA was detected by Western blot. ## p < 0.01 compared with A2780 group.

Article Snippet: Annexin A1 , ABclonal , A1118 , 1:1000.

Techniques: Expressing, Immunohistochemistry, Western Blot

Journal: Cancer Reports

Article Title: ANXA1 / FPR Signaling Contribute to Paclitaxel Resistance in Ovarian Cancer Through PI3K / AKT ‐Associated PGC1α Regulation

doi: 10.1002/cnr2.70637

Figure Lengend Snippet: Inhibition of ANXA1 increases the apoptotic sensitivity of ovarian cancer cells to Paclitaxel. (A) The activity of A2780 cells treated with paclitaxel after the addition of AC2‐26 was detected by MTT. (B) Western Blot analysis of cleaved‐caspase3, Bax, and Bcl2 protein expression in A2780 cells treated with paclitaxel after AC2‐26 addition. In addition, quantification analysis of Cleaved‐caspase3, Bax, and Bcl2 protein and Bax/Bcl2 ratio are shown on the right. (C) The apoptosis rate of A2780 cells treated with paclitaxel after adding AC2‐26 was detected by Annexin V‐FITC/PI double staining. The right figure shows the quantitative analysis of the results. (AC2‐26:3 μm, PA: 0.00625 μg/mL, * p < 0.05 compared with PA group, ** p < 0.01 compared with PA group). (D) The activity of A2780/PA cells treated with paclitaxel after the addition of Boc2 was detected by MTT. (E) Western Blot analysis of cleaved‐caspase3, Bax, and Bcl2 protein expression in A2780/PA cells treated with paclitaxel after Boc2 addition. Quantification analysis of Cleaved‐caspase3, Bax, and Bcl2 protein and Bax/Bcl2 ratio are shown on the right. (F) The apoptosis rate of A2780/PA cells treated with paclitaxel after adding Boc2 was detected by Annexin V‐FITC/PI double staining. The right figure shows the quantitative analysis of the results. (Boc2:10 μm, PA: 10 μg/mL, * p < 0.05 vs. PA group, ** p < 0.01 compared with PA group).

Article Snippet: Annexin A1 , ABclonal , A1118 , 1:1000.

Techniques: Inhibition, Activity Assay, Western Blot, Expressing, Double Staining

Journal: Cancer Reports

Article Title: ANXA1 / FPR Signaling Contribute to Paclitaxel Resistance in Ovarian Cancer Through PI3K / AKT ‐Associated PGC1α Regulation

doi: 10.1002/cnr2.70637

Figure Lengend Snippet: ANXA1 can promote the activation of PI3K/AKT signaling pathway. (A) The STRING database was used to predict ANXA1 related proteins. (B) The mRNA expression of PIK3R1 in A2780 and A2780/PA was detected by qPCR assay, ## p <0.01vs.A2780 group. (C) The protein expressions of PI3K, P‐PI3K, P‐Akt and AKT in A2780 cells treated with paclitaxel after AC2‐26 addition was detected by Western Blot. The figure on the right shows the quantitative analysis of protein expression as P‐Akt/AKT ratio. (AC2‐26:3 μm, PA: 0.00625 μg/mL, ** p < 0.01 compared with PA group). (D) The protein expressions of PI3K, P‐PI3K, P‐Akt, and AKT in A2780/PA cells treated with paclitaxel were detected by Western Blot. The figure on the right is the quantitative analysis of protein expression and P‐Akt/AKT ratio. (Boc2:10 μm, PA: 10 μg/mL, * p < 0.05 vs. PA group, ** p < 0.01 vs. PA group).

Article Snippet: Annexin A1 , ABclonal , A1118 , 1:1000.

Techniques: Activation Assay, Expressing, Western Blot

Journal: Cancer Reports

Article Title: ANXA1 / FPR Signaling Contribute to Paclitaxel Resistance in Ovarian Cancer Through PI3K / AKT ‐Associated PGC1α Regulation

doi: 10.1002/cnr2.70637

Figure Lengend Snippet: ANXA1 can promote PGC1α‐dependent mitochondrial biogenesis. (A) The protein expression levels of PGC1α and downstream related proteins NRF1 and NRF2 in A2780 cells and A2780/PA cells， ## p <0.01campared with A2780 group. The quantitative analysis was shown on the right. (B) The detection of the effect of AC2‐26 on ATP production in A2780 cells treated with paclitaxel. (C) Protein expressions of PGC1α, NRF1, NRF2, and TFAM in A2780 cells treated with paclitaxel were detected by Western Blot. Quantification analysis of protein expression was shown in the right. (AC2‐26:3 μm, PA: 0.00625 μg/mL, * p < 0.05 compared with PA group, ** p < 0.01 compared with PA group).

Article Snippet: Annexin A1 , ABclonal , A1118 , 1:1000.

Techniques: Expressing, Western Blot

Journal: Cancer Reports

Article Title: ANXA1 / FPR Signaling Contribute to Paclitaxel Resistance in Ovarian Cancer Through PI3K / AKT ‐Associated PGC1α Regulation

doi: 10.1002/cnr2.70637

Figure Lengend Snippet: ANXA1 may be involved in the regulation of mitochondrial biogenesis by up‐regulating the PI3K/AKT pathway. (A) The protein expressions of PGC1α, NRF1, NRF2, and TFAM in A2780/PA cells were detected by Western Blot after the addition of Boc2 or PKI‐402 to paclitaxel. (B) Quantitative analysis of protein expression. (C) The detection the effect of Boc2 or PKI‐402 on ATP production in A2780/PA cells treated with paclitaxel. (Boc2:10 μm, PKI‐402:10 μm, PA:10 μg/mL, * p < 0.05 compared with PA group, ** p < 0.01 compared with PA group).

Article Snippet: Annexin A1 , ABclonal , A1118 , 1:1000.

Techniques: Western Blot, Expressing

Journal: Cancer Reports

Article Title: ANXA1 / FPR Signaling Contribute to Paclitaxel Resistance in Ovarian Cancer Through PI3K / AKT ‐Associated PGC1α Regulation

doi: 10.1002/cnr2.70637

Figure Lengend Snippet: Inhibition of ANXA1/FPR signaling suppresses the growth of ovarian cancer cells in vivo. C3H SCID female mice (6 weeks old; 3 per group) were inoculated subcutaneously with A2780/PA cells (3 × 106 per animal). (A–C) Tumor weight and volumes and images of excised tumors from C3H SCID mice at 15 days. (D) Tumor volumes of excised tumors from C3H SCID mice from 0 to 15 days. (E, F) The protein expressions of PI3K, P‐PI3K, AKT, p‐AKT, PGC1α, NRF1, NRF2, and TFAM in Tumor tissues were detected by Western Blot. The quantitative analysis was shown on the right. Tumor volumes were measured as indicated. Data points are presented as mean volume ± s.d. * p < 0.05. ** p < 0.01.

Article Snippet: Annexin A1 , ABclonal , A1118 , 1:1000.

Techniques: Inhibition, In Vivo, Western Blot

Journal: JCI Insight

Article Title: Excess muscle plasma membrane leak disrupts ECM content and shifts macrophage-mediated muscle repair

doi: 10.1172/jci.insight.205137

Figure Lengend Snippet: ( A ) A comparison of proteomic profiles from the CHAPS/NaCl-soluble cellular fraction of WT, Dysf , and DysfA6 muscle shows expansion of ECM-associated proteins ( n = 6–7 mice per genotype). ( B ) Differentially expressed proteins between DysfA6 and WT in the CHAPS/NaCl-soluble cellular and guanidine-HCl–soluble (Gnd-HCl-soluble) ECM fractions of the muscle. ( C and D ) ANXA1 and ANXA2 were increased in both the CHAPS/NaCl- and Gnd-HCl–soluble fractions of DysfA6 compared with Dysf and WT ( n = 6–7 mice per genotype). ( E ) Immunoblot of whole muscle lysates for ANXA1 and ANXA2 confirmed greater expression in the presence of defective repair. ( F and G ) Muscle sections were decellularized to generate myoscaffolds and visualize ECM-associated proteins. Increased ANXA1 and ANXA2 deposition was seen in decellularized myoscaffolds of DysfA6 compared with WT and Dysf by IFM. ** P < 0.01; *** P < 0.005; **** P < 0.001 by 1-way ANOVA followed by Tukey’s multiple comparisons test. Scale bars: 100 μm.

Article Snippet: After 24 hours, 1 mg/mL recombinant ANXA1 (230-00601-50, RayBiotech) and ANXA2 (230-30023-50, RayBiotech) were diluted to 5 μg/mL in macrophage culture media and subsequently added to plated macrophages for 3 hours.

Techniques: Comparison, Western Blot, Expressing

Journal: JCI Insight

Article Title: Excess muscle plasma membrane leak disrupts ECM content and shifts macrophage-mediated muscle repair

doi: 10.1172/jci.insight.205137

Figure Lengend Snippet: ( A ) Muscle cryosections (25 μm) were decellularized, incubated with WT macrophages, and imaged after 24 hours. Z -stack images were acquired and the percentage of macrophages localized on the ECM relative to the total macrophages on the ECM and glass was calculated. More macrophages (F4/80, red) accumulated on Dysf and DysfA6 myoscaffolds compared with WT ( n = 3 mice per genotype denoted by color, 2 myoscaffolds per mouse). Macrophages were derived from 3 WT mice, denoted by symbol shapes. ( B ) WT macrophages exposed to Dysf and DysfA6 myoscaffolds adopted a distinct morphology compared with WT myoscaffolds, with greater increased circularity (arrowhead) induced by Dysf and DysfA6 ( n = 3 mice per genotype, 2 myoscaffolds per animal, and 3 images per myoscaffold). Full arrows indicate macrophages with more elongated morphology. ( C ) Recombinant ANX1 (rANXA1), rANXA2, or combined treatment of macrophages elicited Csf1 expression (macrophages isolated from 3 mice, denoted by symbols, with technical replicates). ( D ) DysfA6 receptor-ligand gene expression analysis with CellChat revealed that Mertk + Trem2 + macrophages signal to FAPs via GAS6, while FAPs signal to macrophages via PROS1 and GAS6. ( E ) Prolonged sarcolemmal leak due to impaired membrane repair increased ANXA1 and ANXA2 ECM deposition, recruiting macrophages to the area of leak and promoting macrophage-FAP crosstalk. All the experiments were conducted 3 independent times. ** P < 0.01; *** P < 0.005; **** P < 0.001 by 1-way ANOVA followed by Tukey’s multiple comparisons test. Scale bars: 100 μm ( A ) and 50 μm ( B ).

Article Snippet: After 24 hours, 1 mg/mL recombinant ANXA1 (230-00601-50, RayBiotech) and ANXA2 (230-30023-50, RayBiotech) were diluted to 5 μg/mL in macrophage culture media and subsequently added to plated macrophages for 3 hours.

Techniques: Incubation, Derivative Assay, Recombinant, Expressing, Isolation, Gene Expression, Membrane

Journal: The FASEB Journal

Article Title: A Novel Dysferlin‐Binding Kinase CK2α Promotes Plasma Membrane Repair in Dysferlinopathy

doi: 10.1096/fj.202500773RRR

Figure Lengend Snippet: Phosphorylation of annexin A1 is decreased in human myocytes with dysferlin mutations. (A) immunoblots of annexin A1, Dysferlin and Vinculin in human myoblasts and myotubes. Line 107 and 379 have dysferlin mutations. (B) The bands of annexinA1 were normalized to those of vinculin and semiquantified. n = 3, Tukey–Kramer test. n.s., Not significant. (C) PhosTag of annexin A1 in human myoblasts. The black line represents the phosphorylation band.

Article Snippet: The antibodies used for immunoblotting and immunoprecipitation were as follows: HA tag (51064–2‐AP; Proteintech, Rosemont, IL), Halo tag (G9281; Promega), dysferlin (NCL‐Hamlet, Leica, Wetzlar, Germany), CK2α/α′ (MCA3031Z; Bio‐Rad), CK2β (ab76025; Abcam, Cambridge, UK), annexin A1 (21990–1‐AP, Proteintech) (#610067, BD Bioscience, Franklinlakes, NJ), pAkt (S127) (ab133458; Abcam), Akt (#9272; CST, Danvers, MA), GFP (M048‐3; MBL, Tokyo, Japan), GAPDH (#2118; CST), Vinculin (#13901; CST), and HRP‐conjugated secondary antibodies (mouse: #7076; CST, rabbit: #7074; CST).

Techniques: Phospho-proteomics, Western Blot

Journal: The FASEB Journal

Article Title: A Novel Dysferlin‐Binding Kinase CK2α Promotes Plasma Membrane Repair in Dysferlinopathy

doi: 10.1096/fj.202500773RRR

Figure Lengend Snippet: Identified annexin A1 as a candidate substrate for CK2α. (A) Peptides were recovered from wild‐type and CK2α KO C2C12 myoblast cells, followed by enrichment of phosphorylated peptides with titanium beads and subsequent mass spectrometry analysis. (B) The MA plot was constructed with log2 ([CK2α KO]/[WT]) on the horizontal axis and the mean amount of detected peptides on the vertical axis. Phosphorylated peptides showing a significant decrease with CK2α KO were identified as those decreasing below −1.0 on the horizontal axis (indicated by the red line). (C) A list of phosphorylated peptides detected in this study, which are reported to be involved in cell membrane repair. (D) With a Qual Browser, the detection intensity of the phosphorylated peptides of annexin A1 was compared between the WT and CK2α KO strains. (E) PhosTag was utilized to assess the phosphorylation of annexin A1 in WT and CK2α KO C2C12 myoblasts. The black line represents the phosphorylation band observed in the WT.

Article Snippet: The antibodies used for immunoblotting and immunoprecipitation were as follows: HA tag (51064–2‐AP; Proteintech, Rosemont, IL), Halo tag (G9281; Promega), dysferlin (NCL‐Hamlet, Leica, Wetzlar, Germany), CK2α/α′ (MCA3031Z; Bio‐Rad), CK2β (ab76025; Abcam, Cambridge, UK), annexin A1 (21990–1‐AP, Proteintech) (#610067, BD Bioscience, Franklinlakes, NJ), pAkt (S127) (ab133458; Abcam), Akt (#9272; CST, Danvers, MA), GFP (M048‐3; MBL, Tokyo, Japan), GAPDH (#2118; CST), Vinculin (#13901; CST), and HRP‐conjugated secondary antibodies (mouse: #7076; CST, rabbit: #7074; CST).

Techniques: Mass Spectrometry, Construct, Membrane, Phospho-proteomics

Journal: The FASEB Journal

Article Title: A Novel Dysferlin‐Binding Kinase CK2α Promotes Plasma Membrane Repair in Dysferlinopathy

doi: 10.1096/fj.202500773RRR

Figure Lengend Snippet: Schematic diagram of plasma membrane repair mediated by dysferlin, as elucidated in this study. (A) At steady state, dysferlin spans the membrane at the C‐terminus, and CK2 binds to dysferlin at region II. (B) When the cell membrane is injured, extracellular calcium ions influx, leading to the accumulation of dysferlin at the injury site, where it is believed to bind to the cytoplasmic side of the membrane via its C2 domain. In the presence of dysferlin, CK2 also efficiently distributes to the site of membrane injury, maintaining its kinase activity by interacting with dysferlin. Phosphorylated annexin A1 recruits intracellular vesicles to the injury site. (C) The mobilized intracellular vesicles are hypothesized to serve as a source of phospholipids, which act as material for the patch that seals the damaged site in cellular membrane repair.

Article Snippet: The antibodies used for immunoblotting and immunoprecipitation were as follows: HA tag (51064–2‐AP; Proteintech, Rosemont, IL), Halo tag (G9281; Promega), dysferlin (NCL‐Hamlet, Leica, Wetzlar, Germany), CK2α/α′ (MCA3031Z; Bio‐Rad), CK2β (ab76025; Abcam, Cambridge, UK), annexin A1 (21990–1‐AP, Proteintech) (#610067, BD Bioscience, Franklinlakes, NJ), pAkt (S127) (ab133458; Abcam), Akt (#9272; CST, Danvers, MA), GFP (M048‐3; MBL, Tokyo, Japan), GAPDH (#2118; CST), Vinculin (#13901; CST), and HRP‐conjugated secondary antibodies (mouse: #7076; CST, rabbit: #7074; CST).

Techniques: Clinical Proteomics, Membrane, Activity Assay

Journal: bioRxiv

Article Title: Transplanting ANXA1⁻ CD8⁺ Naïve T cells Delay Aging Through Senolysis

doi: 10.64898/2026.02.21.707223

Figure Lengend Snippet: (A) Gene Set Enrichment Analysis (GSEA) comparing CD8⁺ naïve T cells from Elderly versus other groups of donors. Plots show significant enrichment of the Cellular Senescence (GO:0090398) and Senescence-Associated Secretory Phenotype (SASP; R-HSA-2559582) gene sets in the Elderly group. NES, normalized enrichment score. (B) Heatmap displaying the expression of senescence-associated signature genes in CD8⁺ naïve T cells across all donors. Each row is scaled using a Z-score. (C) Violin plot showing the normalized expression level of ANXA1 in CD8⁺ naïve T cells across the five age groups from scRNA-seq data. (D) Quantification of ANXA1 surface expression in CD8⁺ naïve T cells across the five age groups, as measured by flow cytometry. Each point represents an individual donor (Kid, n = 31; Young, n = 98; Middle-aged, n = 93; Pre-Elderly, n = 52; Elderly, n = 63). (E) Representative flowcytometry plots and summary quantification of total ANXA1 expression levels in CD8⁺ naïve T cells from human PBMCs (n = 5 per group). (F) Pathway enrichment analysis comparing human ANXA1⁺ versus ANXA1⁻ CD8⁺ naïve T cells from RNA-seq. The bubble plot shows key pathways significantly altered between the two subsets. Bubble size corresponds to the number of genes, and color indicates enrichment significance. (G) Representative immunofluorescence images for ANXA1 and p16 INK4a expression in human CD8⁺ naïve T cells. Scale bar, 10 µm. (H) Representative flow cytometry histogram and quantification p16 INK4a expression in human ANXA1⁺ versus ANXA1⁻ CD8⁺ naïve T cells (n = 5 per group). (I) Representative histogram of Cell Trace Violet (CTV) dye dilution in human ANXA1⁻ and ANXA1⁺ CD8⁺ naïve T cells after 72 hours of stimulation. Data are presented as mean ± SEM. P -value was determined by one-way ANOVA and two-tailed unpaired t-test.

Article Snippet: The following antibodies and reagents were used for human and mouse flow cytometry: APC anti-human Annexin A1[Clone: 74/3] (BioLegend, cat: 831604), PE anti-human RANTES (CCL5) [Clone: VL1] (BioLegend, cat: 515503), PerCP/Cy5.5 anti-human CD45RA [Clone: HI100] (BioLegend, cat: 304122), Brilliant Violet® 605 anti-human CD62L [Clone: DREG-56] (BioLegend, cat: 304834), PE (Phycoerythrin)/Cy7® anti-human CD25 [Clone: BC96] (BioLegend, cat: 302611), APC anti-human CD45 [clone: HI30] (BioLegend, cat: 304012), PE anti-human CD3 [Clone: UCHT1] (BioLegend, cat: 300441), FITC anti-human CD8a [Clone: RPA-T8] (BioLegend, cat: 301050), anti-CDKN2A/p16 INK4a antibody (abcam, cat. ab189034), DAPI (BD Pharmingen, cat: 564907), PE (Phycoerythrin)/Cy7® anti-mouse CD69 [clone: H1.2F3] (BioLegend, cat: 104512), PE anti-mouse CD25 [Clone: PC61] (BioLegend, cat: 102008), Brilliant Violet 421 anti-mouse CD25 [Clone: PC61] (BD Pharmingen, cat: 562606), Alexa Fluor® 700 anti-mouse CD45.2 [Clone: 104] (BioLegend, cat: 109822), PE anti-mouse CD45.1 [Clone: A20] (BioLegend, cat: 110708), PE-Cy7 anti-mouse IFN-γ [Clone: XMG1.2] (BD Pharmingen, cat: 557649), APC/Cyanine7 anti-mouse CD279 (PD-1) [Clone: 29F.1A12] (BioLegend, cat: 135223), Brilliant Violet 650 anti-mouse CD223 (LAG-3) [Clone: C9B7W] (BioLegend, cat: 125227), Annexin A1 Monoclonal antibody [Clone: 1E1B7] (Proteintech, cat: 66344-1-IG), Alexa Fluor® 488 Anti-Annexin A1/ANXA1 antibody [Clone: EPR19342] (abcam, cat: ab225513), PE anti-mouse/human CD44 [Clone: IM7] (BioLegend, cat: 103024), APC anti-mouse CD62L [Clone: MEL-14] (BioLegend, cat: 104412), PE Anti-Mouse CD107a/LAMP-1 Antibody [Clone: 1D4B] (Elabscience, cat: E-AB-F1254D), PE-CF594 Hamster Anti-Mouse CD3e [Clone: 145-2C11] (BD Pharmingen, cat: 562286), PerCP/Cyanine5.5 anti-mouse CD8a [Clone: 53-6.7] (BioLegend, cat: 100734).

Techniques: Expressing, Flow Cytometry, RNA Sequencing, Immunofluorescence, Two Tailed Test

Journal: bioRxiv

Article Title: Transplanting ANXA1⁻ CD8⁺ Naïve T cells Delay Aging Through Senolysis

doi: 10.64898/2026.02.21.707223

Figure Lengend Snippet: (A) Representative flow cytometric analysis of CCL5 expression on CD8 + naïve T cells in samples of young (19-35 y) and old (>65 y) group (n= 5 per group). (B) Sub-clustering of CD8⁺ naïve T cells from the scRNA-seq dataset. UMAP projection showing distinct subclusters (C1-C6) identified within the CD8⁺ naïve T cell population. (C) Feature plot overlaying normalized ANXA1 expression onto the UMAP projection from ( B ), highlighting its specific enrichment in the C4 subcluster. (D) Scatter plots showing the correlation between the relative abundance of each subcluster (as a percentage of total CD8⁺ naïve T cells) and donor age. Lines represent linear regression fits; Pearson correlation coefficient (R) and P -values are indicated. ( E and F ) Violin plots ( E ) and heatmap ( F ) validating the naïve identity of both ANXA1 − and ANXA1⁺ CD8⁺ naïve T cells defined in human scRNA-seq data. Average expression of canonical lineage markers for naïve, memory, effector, and exhausted T cells is compared across three populations with CD8⁺ effector memory (Tem) cells as a non-naïve control. Expression values are scaled per row. (G) Representative immunofluorescence images and quantification of ANXA1 surface expression on mouse CD8⁺ naïve T cells from Young (8-week-old, n = 5) and Old (13-month-old, n = 5) mice. Scale bar, 5 µm. (H) Representative flow cytometry results and quantifications on the percentage of CD8 + naïve T cells and surface ANXA1 expression in young and old mice (n = 3 per group). Data in quantification plots are presented as mean ± SEM. Statistical significance was determined by one-way ANOVA and two-tailed unpaired t-test.

Article Snippet: The following antibodies and reagents were used for human and mouse flow cytometry: APC anti-human Annexin A1[Clone: 74/3] (BioLegend, cat: 831604), PE anti-human RANTES (CCL5) [Clone: VL1] (BioLegend, cat: 515503), PerCP/Cy5.5 anti-human CD45RA [Clone: HI100] (BioLegend, cat: 304122), Brilliant Violet® 605 anti-human CD62L [Clone: DREG-56] (BioLegend, cat: 304834), PE (Phycoerythrin)/Cy7® anti-human CD25 [Clone: BC96] (BioLegend, cat: 302611), APC anti-human CD45 [clone: HI30] (BioLegend, cat: 304012), PE anti-human CD3 [Clone: UCHT1] (BioLegend, cat: 300441), FITC anti-human CD8a [Clone: RPA-T8] (BioLegend, cat: 301050), anti-CDKN2A/p16 INK4a antibody (abcam, cat. ab189034), DAPI (BD Pharmingen, cat: 564907), PE (Phycoerythrin)/Cy7® anti-mouse CD69 [clone: H1.2F3] (BioLegend, cat: 104512), PE anti-mouse CD25 [Clone: PC61] (BioLegend, cat: 102008), Brilliant Violet 421 anti-mouse CD25 [Clone: PC61] (BD Pharmingen, cat: 562606), Alexa Fluor® 700 anti-mouse CD45.2 [Clone: 104] (BioLegend, cat: 109822), PE anti-mouse CD45.1 [Clone: A20] (BioLegend, cat: 110708), PE-Cy7 anti-mouse IFN-γ [Clone: XMG1.2] (BD Pharmingen, cat: 557649), APC/Cyanine7 anti-mouse CD279 (PD-1) [Clone: 29F.1A12] (BioLegend, cat: 135223), Brilliant Violet 650 anti-mouse CD223 (LAG-3) [Clone: C9B7W] (BioLegend, cat: 125227), Annexin A1 Monoclonal antibody [Clone: 1E1B7] (Proteintech, cat: 66344-1-IG), Alexa Fluor® 488 Anti-Annexin A1/ANXA1 antibody [Clone: EPR19342] (abcam, cat: ab225513), PE anti-mouse/human CD44 [Clone: IM7] (BioLegend, cat: 103024), APC anti-mouse CD62L [Clone: MEL-14] (BioLegend, cat: 104412), PE Anti-Mouse CD107a/LAMP-1 Antibody [Clone: 1D4B] (Elabscience, cat: E-AB-F1254D), PE-CF594 Hamster Anti-Mouse CD3e [Clone: 145-2C11] (BD Pharmingen, cat: 562286), PerCP/Cyanine5.5 anti-mouse CD8a [Clone: 53-6.7] (BioLegend, cat: 100734).

Techniques: Expressing, Control, Immunofluorescence, Flow Cytometry, Two Tailed Test

Journal: bioRxiv

Article Title: Transplanting ANXA1⁻ CD8⁺ Naïve T cells Delay Aging Through Senolysis

doi: 10.64898/2026.02.21.707223

Figure Lengend Snippet: (A) Heatmap of DEGs from RNA-seq of resting WT and Anxa1 KO CD8⁺ naïve T cells. (B) GO enrichment bubble plot of pathways upregulated in Anxa1 KO versus WT CD8⁺ naïve T cells at basal state.

Article Snippet: The following antibodies and reagents were used for human and mouse flow cytometry: APC anti-human Annexin A1[Clone: 74/3] (BioLegend, cat: 831604), PE anti-human RANTES (CCL5) [Clone: VL1] (BioLegend, cat: 515503), PerCP/Cy5.5 anti-human CD45RA [Clone: HI100] (BioLegend, cat: 304122), Brilliant Violet® 605 anti-human CD62L [Clone: DREG-56] (BioLegend, cat: 304834), PE (Phycoerythrin)/Cy7® anti-human CD25 [Clone: BC96] (BioLegend, cat: 302611), APC anti-human CD45 [clone: HI30] (BioLegend, cat: 304012), PE anti-human CD3 [Clone: UCHT1] (BioLegend, cat: 300441), FITC anti-human CD8a [Clone: RPA-T8] (BioLegend, cat: 301050), anti-CDKN2A/p16 INK4a antibody (abcam, cat. ab189034), DAPI (BD Pharmingen, cat: 564907), PE (Phycoerythrin)/Cy7® anti-mouse CD69 [clone: H1.2F3] (BioLegend, cat: 104512), PE anti-mouse CD25 [Clone: PC61] (BioLegend, cat: 102008), Brilliant Violet 421 anti-mouse CD25 [Clone: PC61] (BD Pharmingen, cat: 562606), Alexa Fluor® 700 anti-mouse CD45.2 [Clone: 104] (BioLegend, cat: 109822), PE anti-mouse CD45.1 [Clone: A20] (BioLegend, cat: 110708), PE-Cy7 anti-mouse IFN-γ [Clone: XMG1.2] (BD Pharmingen, cat: 557649), APC/Cyanine7 anti-mouse CD279 (PD-1) [Clone: 29F.1A12] (BioLegend, cat: 135223), Brilliant Violet 650 anti-mouse CD223 (LAG-3) [Clone: C9B7W] (BioLegend, cat: 125227), Annexin A1 Monoclonal antibody [Clone: 1E1B7] (Proteintech, cat: 66344-1-IG), Alexa Fluor® 488 Anti-Annexin A1/ANXA1 antibody [Clone: EPR19342] (abcam, cat: ab225513), PE anti-mouse/human CD44 [Clone: IM7] (BioLegend, cat: 103024), APC anti-mouse CD62L [Clone: MEL-14] (BioLegend, cat: 104412), PE Anti-Mouse CD107a/LAMP-1 Antibody [Clone: 1D4B] (Elabscience, cat: E-AB-F1254D), PE-CF594 Hamster Anti-Mouse CD3e [Clone: 145-2C11] (BD Pharmingen, cat: 562286), PerCP/Cyanine5.5 anti-mouse CD8a [Clone: 53-6.7] (BioLegend, cat: 100734).

Techniques: RNA Sequencing

Journal: bioRxiv

Article Title: Transplanting ANXA1⁻ CD8⁺ Naïve T cells Delay Aging Through Senolysis

doi: 10.64898/2026.02.21.707223

Figure Lengend Snippet: (A) Heatmap of DEGs in activated CD8⁺ naïve T cells from WT and Anxa1 KO mice. Cells were stimulated with anti-CD3/CD28 antibodies for 12 hours prior to RNA-seq. Each row is scaled using a Z-score. (B) GO enrichment of pathways upregulated in activated Anxa1 KO CD8⁺ naïve T cells compared with WT. ( C and D ) Representative flow cytometry histograms and quantification of activation markers CD69 ( C ) and CD25 ( D ) on WT and Anxa1 KO CD8⁺ naïve T cells 12 hours post-stimulation (n = 6 per group). (E) Representative flow cytometry plots and quantification of IFNγ-producing cells within WT and Anxa1 KO CD8⁺ T cells after stimulation (n = 6 per group). (F) Representative flow cytometry plots and quantification of the degranulation marker CD107a on WT and Anxa1 KO CD8⁺ T cells after stimulation (n = 6 per group). (G) Schematic of the B16-F10 inoculation model. C57BL/6 mice were inoculated subcutaneously with B16-F10 melanoma cells, followed by adoptive transfer of CD8⁺ T cells isolated from either WT or Anxa1 KO mice. (H) Representative image and quantification of excised tumors at day 20 post-inoculation. (n = 6 per group). Data in quantification plots are presented as mean ± SEM. P-values were determined by two-tailed unpaired t-test.

Article Snippet: The following antibodies and reagents were used for human and mouse flow cytometry: APC anti-human Annexin A1[Clone: 74/3] (BioLegend, cat: 831604), PE anti-human RANTES (CCL5) [Clone: VL1] (BioLegend, cat: 515503), PerCP/Cy5.5 anti-human CD45RA [Clone: HI100] (BioLegend, cat: 304122), Brilliant Violet® 605 anti-human CD62L [Clone: DREG-56] (BioLegend, cat: 304834), PE (Phycoerythrin)/Cy7® anti-human CD25 [Clone: BC96] (BioLegend, cat: 302611), APC anti-human CD45 [clone: HI30] (BioLegend, cat: 304012), PE anti-human CD3 [Clone: UCHT1] (BioLegend, cat: 300441), FITC anti-human CD8a [Clone: RPA-T8] (BioLegend, cat: 301050), anti-CDKN2A/p16 INK4a antibody (abcam, cat. ab189034), DAPI (BD Pharmingen, cat: 564907), PE (Phycoerythrin)/Cy7® anti-mouse CD69 [clone: H1.2F3] (BioLegend, cat: 104512), PE anti-mouse CD25 [Clone: PC61] (BioLegend, cat: 102008), Brilliant Violet 421 anti-mouse CD25 [Clone: PC61] (BD Pharmingen, cat: 562606), Alexa Fluor® 700 anti-mouse CD45.2 [Clone: 104] (BioLegend, cat: 109822), PE anti-mouse CD45.1 [Clone: A20] (BioLegend, cat: 110708), PE-Cy7 anti-mouse IFN-γ [Clone: XMG1.2] (BD Pharmingen, cat: 557649), APC/Cyanine7 anti-mouse CD279 (PD-1) [Clone: 29F.1A12] (BioLegend, cat: 135223), Brilliant Violet 650 anti-mouse CD223 (LAG-3) [Clone: C9B7W] (BioLegend, cat: 125227), Annexin A1 Monoclonal antibody [Clone: 1E1B7] (Proteintech, cat: 66344-1-IG), Alexa Fluor® 488 Anti-Annexin A1/ANXA1 antibody [Clone: EPR19342] (abcam, cat: ab225513), PE anti-mouse/human CD44 [Clone: IM7] (BioLegend, cat: 103024), APC anti-mouse CD62L [Clone: MEL-14] (BioLegend, cat: 104412), PE Anti-Mouse CD107a/LAMP-1 Antibody [Clone: 1D4B] (Elabscience, cat: E-AB-F1254D), PE-CF594 Hamster Anti-Mouse CD3e [Clone: 145-2C11] (BD Pharmingen, cat: 562286), PerCP/Cyanine5.5 anti-mouse CD8a [Clone: 53-6.7] (BioLegend, cat: 100734).

Techniques: RNA Sequencing, Flow Cytometry, Activation Assay, Marker, Adoptive Transfer Assay, Isolation, Two Tailed Test

Journal: bioRxiv

Article Title: Transplanting ANXA1⁻ CD8⁺ Naïve T cells Delay Aging Through Senolysis

doi: 10.64898/2026.02.21.707223

Figure Lengend Snippet: (A) Representative flow cytometry histograms and quantifications of CTV-staining, CD69 and CD25 expression of ANXA1 − CD8 + naïve T cells from young and aged mice (n = 3 per group). (B) Representative flow cytometry results and quantifications of Tc1 differentiation of ANXA1 − CD8 + naïve T cells from young and aged mice. (C) Representative images and quantification from a transwell invasion assay showing that both young and old mouse ANXA1⁻ T cells, but not ANXA1⁺ T cells (n = 5 per group), significantly inhibit the invasion of U-2 OS tumor cells, with arrows indicate tumor cells passed the transwell membrane. Data are presented as mean ± SEM. P -values were determined by one-way ANOVA and two-tailed unpaired t-test.

Article Snippet: The following antibodies and reagents were used for human and mouse flow cytometry: APC anti-human Annexin A1[Clone: 74/3] (BioLegend, cat: 831604), PE anti-human RANTES (CCL5) [Clone: VL1] (BioLegend, cat: 515503), PerCP/Cy5.5 anti-human CD45RA [Clone: HI100] (BioLegend, cat: 304122), Brilliant Violet® 605 anti-human CD62L [Clone: DREG-56] (BioLegend, cat: 304834), PE (Phycoerythrin)/Cy7® anti-human CD25 [Clone: BC96] (BioLegend, cat: 302611), APC anti-human CD45 [clone: HI30] (BioLegend, cat: 304012), PE anti-human CD3 [Clone: UCHT1] (BioLegend, cat: 300441), FITC anti-human CD8a [Clone: RPA-T8] (BioLegend, cat: 301050), anti-CDKN2A/p16 INK4a antibody (abcam, cat. ab189034), DAPI (BD Pharmingen, cat: 564907), PE (Phycoerythrin)/Cy7® anti-mouse CD69 [clone: H1.2F3] (BioLegend, cat: 104512), PE anti-mouse CD25 [Clone: PC61] (BioLegend, cat: 102008), Brilliant Violet 421 anti-mouse CD25 [Clone: PC61] (BD Pharmingen, cat: 562606), Alexa Fluor® 700 anti-mouse CD45.2 [Clone: 104] (BioLegend, cat: 109822), PE anti-mouse CD45.1 [Clone: A20] (BioLegend, cat: 110708), PE-Cy7 anti-mouse IFN-γ [Clone: XMG1.2] (BD Pharmingen, cat: 557649), APC/Cyanine7 anti-mouse CD279 (PD-1) [Clone: 29F.1A12] (BioLegend, cat: 135223), Brilliant Violet 650 anti-mouse CD223 (LAG-3) [Clone: C9B7W] (BioLegend, cat: 125227), Annexin A1 Monoclonal antibody [Clone: 1E1B7] (Proteintech, cat: 66344-1-IG), Alexa Fluor® 488 Anti-Annexin A1/ANXA1 antibody [Clone: EPR19342] (abcam, cat: ab225513), PE anti-mouse/human CD44 [Clone: IM7] (BioLegend, cat: 103024), APC anti-mouse CD62L [Clone: MEL-14] (BioLegend, cat: 104412), PE Anti-Mouse CD107a/LAMP-1 Antibody [Clone: 1D4B] (Elabscience, cat: E-AB-F1254D), PE-CF594 Hamster Anti-Mouse CD3e [Clone: 145-2C11] (BD Pharmingen, cat: 562286), PerCP/Cyanine5.5 anti-mouse CD8a [Clone: 53-6.7] (BioLegend, cat: 100734).

Techniques: Flow Cytometry, Staining, Expressing, Transwell Invasion Assay, Membrane, Two Tailed Test

Journal: bioRxiv

Article Title: Transplanting ANXA1⁻ CD8⁺ Naïve T cells Delay Aging Through Senolysis

doi: 10.64898/2026.02.21.707223

Figure Lengend Snippet: (A) Schematic of the co-culture experiment using fibroblasts engineered to express p16 INK4a -GFP as a model of senescent cells. Human ANXA1 − or ANXA1 + CD8 + naïve T cells using FACS were co-cultured with fibroblasts for live imaging. (B) Representative stills from live-cell imaging showing the targeting and clearance of p16 INK4a -GFP⁺ senescent cells by ANXA1 − or ANXA1 + CD8 + naïve T cells 6 hours post-culture. (C) Representative immunofluorescence images and quantification of p53- and γH2AX- expressing fibroblasts after co-culture in UV-B induced senescence model (n = 6 per group), corresponding with . (D) Schematic of an Etoposide-induced senescence model. (E) Representative immunofluorescence images and quantification showing a significant reduction of p16 INK4a -positive senescent fibroblasts after co-culture with ANXA1⁻ T cells, but not with ANXA1⁺ T cells (n = 5 per group). Scale bar, 10 µm. (F) Representative immunofluorescence images and quantification of GZMB-secreting cells from different T cell groups after co-culture in (E) (n = 5 per group). Scale bar, 10 µm. Data are presented as mean ± SEM. P -values were determined by one-way ANOVA and two-tailed unpaired t-test.

Article Snippet: The following antibodies and reagents were used for human and mouse flow cytometry: APC anti-human Annexin A1[Clone: 74/3] (BioLegend, cat: 831604), PE anti-human RANTES (CCL5) [Clone: VL1] (BioLegend, cat: 515503), PerCP/Cy5.5 anti-human CD45RA [Clone: HI100] (BioLegend, cat: 304122), Brilliant Violet® 605 anti-human CD62L [Clone: DREG-56] (BioLegend, cat: 304834), PE (Phycoerythrin)/Cy7® anti-human CD25 [Clone: BC96] (BioLegend, cat: 302611), APC anti-human CD45 [clone: HI30] (BioLegend, cat: 304012), PE anti-human CD3 [Clone: UCHT1] (BioLegend, cat: 300441), FITC anti-human CD8a [Clone: RPA-T8] (BioLegend, cat: 301050), anti-CDKN2A/p16 INK4a antibody (abcam, cat. ab189034), DAPI (BD Pharmingen, cat: 564907), PE (Phycoerythrin)/Cy7® anti-mouse CD69 [clone: H1.2F3] (BioLegend, cat: 104512), PE anti-mouse CD25 [Clone: PC61] (BioLegend, cat: 102008), Brilliant Violet 421 anti-mouse CD25 [Clone: PC61] (BD Pharmingen, cat: 562606), Alexa Fluor® 700 anti-mouse CD45.2 [Clone: 104] (BioLegend, cat: 109822), PE anti-mouse CD45.1 [Clone: A20] (BioLegend, cat: 110708), PE-Cy7 anti-mouse IFN-γ [Clone: XMG1.2] (BD Pharmingen, cat: 557649), APC/Cyanine7 anti-mouse CD279 (PD-1) [Clone: 29F.1A12] (BioLegend, cat: 135223), Brilliant Violet 650 anti-mouse CD223 (LAG-3) [Clone: C9B7W] (BioLegend, cat: 125227), Annexin A1 Monoclonal antibody [Clone: 1E1B7] (Proteintech, cat: 66344-1-IG), Alexa Fluor® 488 Anti-Annexin A1/ANXA1 antibody [Clone: EPR19342] (abcam, cat: ab225513), PE anti-mouse/human CD44 [Clone: IM7] (BioLegend, cat: 103024), APC anti-mouse CD62L [Clone: MEL-14] (BioLegend, cat: 104412), PE Anti-Mouse CD107a/LAMP-1 Antibody [Clone: 1D4B] (Elabscience, cat: E-AB-F1254D), PE-CF594 Hamster Anti-Mouse CD3e [Clone: 145-2C11] (BD Pharmingen, cat: 562286), PerCP/Cyanine5.5 anti-mouse CD8a [Clone: 53-6.7] (BioLegend, cat: 100734).

Techniques: Co-Culture Assay, Cell Culture, Imaging, Live Cell Imaging, Immunofluorescence, Expressing, Two Tailed Test

Journal: bioRxiv

Article Title: Transplanting ANXA1⁻ CD8⁺ Naïve T cells Delay Aging Through Senolysis

doi: 10.64898/2026.02.21.707223

Figure Lengend Snippet: (A) Representative stills from live-cell imaging of p16 INK4a -GFP⁺ senescent fibroblasts (green), normal fibroblasts (grey) co-cultured with sorted surface ANXA1⁻ CD8⁺ naïve T cells (magenta), with arrows indicate interaction spots. (B) Schematic diagram of the in vitro UV-B induced senescence co-culture system. Mouse fibroblasts (mFib) were induced into senescence by UV-B radiation and then co-cultured for 5 days with different subsets of sorted CD8⁺ naïve T cells. (C) Representative immunofluorescence images and quantification of p21-positive fibroblasts after co-culture with the indicated T cell subsets (n = 6 per group). Scale bar, 10 µm. (D) Western blot analysis of p16 INK4a protein levels in senescent fibroblasts following co-culture. (E) Schematic of the doxorubicin (Doxo)-induced systemic senescence model. Mice received a single intravenous ( i.v. ) injection of ANXA1⁻ CD8⁺ naïve T cells prior to treatment with Doxo. (F) Rotarod test quantification showing motor coordination and endurance in mice (n = 5 per group). (G) Weight loss of mice in different groups (n = 5 per group) in Doxo-induced senescence model. Initial body weight was normalized to 100%. (H) Representative images and quantifications of p16 INK4a and γH2AX in the heart and kidney from Doxo-treated mice (n = 5 per group). Scale bar, 5 µm. Data in quantification plots are presented as mean ± SEM. P-values were determined by one-way ANOVA and two-tailed unpaired t-test.

Article Snippet: The following antibodies and reagents were used for human and mouse flow cytometry: APC anti-human Annexin A1[Clone: 74/3] (BioLegend, cat: 831604), PE anti-human RANTES (CCL5) [Clone: VL1] (BioLegend, cat: 515503), PerCP/Cy5.5 anti-human CD45RA [Clone: HI100] (BioLegend, cat: 304122), Brilliant Violet® 605 anti-human CD62L [Clone: DREG-56] (BioLegend, cat: 304834), PE (Phycoerythrin)/Cy7® anti-human CD25 [Clone: BC96] (BioLegend, cat: 302611), APC anti-human CD45 [clone: HI30] (BioLegend, cat: 304012), PE anti-human CD3 [Clone: UCHT1] (BioLegend, cat: 300441), FITC anti-human CD8a [Clone: RPA-T8] (BioLegend, cat: 301050), anti-CDKN2A/p16 INK4a antibody (abcam, cat. ab189034), DAPI (BD Pharmingen, cat: 564907), PE (Phycoerythrin)/Cy7® anti-mouse CD69 [clone: H1.2F3] (BioLegend, cat: 104512), PE anti-mouse CD25 [Clone: PC61] (BioLegend, cat: 102008), Brilliant Violet 421 anti-mouse CD25 [Clone: PC61] (BD Pharmingen, cat: 562606), Alexa Fluor® 700 anti-mouse CD45.2 [Clone: 104] (BioLegend, cat: 109822), PE anti-mouse CD45.1 [Clone: A20] (BioLegend, cat: 110708), PE-Cy7 anti-mouse IFN-γ [Clone: XMG1.2] (BD Pharmingen, cat: 557649), APC/Cyanine7 anti-mouse CD279 (PD-1) [Clone: 29F.1A12] (BioLegend, cat: 135223), Brilliant Violet 650 anti-mouse CD223 (LAG-3) [Clone: C9B7W] (BioLegend, cat: 125227), Annexin A1 Monoclonal antibody [Clone: 1E1B7] (Proteintech, cat: 66344-1-IG), Alexa Fluor® 488 Anti-Annexin A1/ANXA1 antibody [Clone: EPR19342] (abcam, cat: ab225513), PE anti-mouse/human CD44 [Clone: IM7] (BioLegend, cat: 103024), APC anti-mouse CD62L [Clone: MEL-14] (BioLegend, cat: 104412), PE Anti-Mouse CD107a/LAMP-1 Antibody [Clone: 1D4B] (Elabscience, cat: E-AB-F1254D), PE-CF594 Hamster Anti-Mouse CD3e [Clone: 145-2C11] (BD Pharmingen, cat: 562286), PerCP/Cyanine5.5 anti-mouse CD8a [Clone: 53-6.7] (BioLegend, cat: 100734).

Techniques: Live Cell Imaging, Cell Culture, In Vitro, Co-Culture Assay, Immunofluorescence, Western Blot, Injection, Two Tailed Test

Journal: bioRxiv

Article Title: Transplanting ANXA1⁻ CD8⁺ Naïve T cells Delay Aging Through Senolysis

doi: 10.64898/2026.02.21.707223

Figure Lengend Snippet: (A) Schematic of the long-term therapeutic study design. Old (13-month-old) mice received monthly intravenous ( i.v. ) transfers of Anxa1⁻ CD8⁺ naïve T cells (Old-Treated group) or a vehicle control (Old-Ctrl group) for the duration of the study. (B) Kaplan-Meier survival curves for all mice with sexes combined. Statistical significance was determined by the Log-rank test. Median survival is indicated by dashed lines. (C) Representative photographs of mice at 30 months of age, highlighting the improved physical condition in the treated mouse. (D) Single-cell t-SNE projections of total CD45⁺ immune cells from the bone marrow of Old-Ctrl and Old-Treated mice (n = 2 per group). Left: Integrated UMAP colored by major cell lineages. Right: The same UMAP split by treatment group to visualize compositional shifts. (E) Box plots show the relative abundance of Inflammatory, Lymphoid, and Myeloid cell clusters as a percentage of total CD45⁺ cells between the two groups. (F) Violin plots comparing the senescence score, calculated per cell, between the Old-Ctrl and Old-Treated groups. The gene list for this score is provided in the Methods section. Statistical significance was determined by two-tailed unpaired t-test. (G) Density plot comparing the “Aging score” for all CD8⁺ T cells from the three experimental groups. The gene list for this score is provided in the Methods section. (H) GSEA enrichment analysis comparing Old-Treated versus Old-Ctrl CD8⁺ T cells. Bar plot shows NES for selected Hallmark gene sets. Purple bars indicate pathways enriched in Old-Ctrl, while orange bars indicate pathways enriched in Old-Treated cells.

Article Snippet: The following antibodies and reagents were used for human and mouse flow cytometry: APC anti-human Annexin A1[Clone: 74/3] (BioLegend, cat: 831604), PE anti-human RANTES (CCL5) [Clone: VL1] (BioLegend, cat: 515503), PerCP/Cy5.5 anti-human CD45RA [Clone: HI100] (BioLegend, cat: 304122), Brilliant Violet® 605 anti-human CD62L [Clone: DREG-56] (BioLegend, cat: 304834), PE (Phycoerythrin)/Cy7® anti-human CD25 [Clone: BC96] (BioLegend, cat: 302611), APC anti-human CD45 [clone: HI30] (BioLegend, cat: 304012), PE anti-human CD3 [Clone: UCHT1] (BioLegend, cat: 300441), FITC anti-human CD8a [Clone: RPA-T8] (BioLegend, cat: 301050), anti-CDKN2A/p16 INK4a antibody (abcam, cat. ab189034), DAPI (BD Pharmingen, cat: 564907), PE (Phycoerythrin)/Cy7® anti-mouse CD69 [clone: H1.2F3] (BioLegend, cat: 104512), PE anti-mouse CD25 [Clone: PC61] (BioLegend, cat: 102008), Brilliant Violet 421 anti-mouse CD25 [Clone: PC61] (BD Pharmingen, cat: 562606), Alexa Fluor® 700 anti-mouse CD45.2 [Clone: 104] (BioLegend, cat: 109822), PE anti-mouse CD45.1 [Clone: A20] (BioLegend, cat: 110708), PE-Cy7 anti-mouse IFN-γ [Clone: XMG1.2] (BD Pharmingen, cat: 557649), APC/Cyanine7 anti-mouse CD279 (PD-1) [Clone: 29F.1A12] (BioLegend, cat: 135223), Brilliant Violet 650 anti-mouse CD223 (LAG-3) [Clone: C9B7W] (BioLegend, cat: 125227), Annexin A1 Monoclonal antibody [Clone: 1E1B7] (Proteintech, cat: 66344-1-IG), Alexa Fluor® 488 Anti-Annexin A1/ANXA1 antibody [Clone: EPR19342] (abcam, cat: ab225513), PE anti-mouse/human CD44 [Clone: IM7] (BioLegend, cat: 103024), APC anti-mouse CD62L [Clone: MEL-14] (BioLegend, cat: 104412), PE Anti-Mouse CD107a/LAMP-1 Antibody [Clone: 1D4B] (Elabscience, cat: E-AB-F1254D), PE-CF594 Hamster Anti-Mouse CD3e [Clone: 145-2C11] (BD Pharmingen, cat: 562286), PerCP/Cyanine5.5 anti-mouse CD8a [Clone: 53-6.7] (BioLegend, cat: 100734).

Techniques: Control, Single Cell, Two Tailed Test