i ap Search Results


antibodies against p p70s6k  (Proteintech)
96
Proteintech antibodies against p p70s6k
Antibodies Against P P70s6k, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 96 stars, based on 1 article reviews
antibodies against p p70s6k - by Bioz Stars, 2026-06
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collagen type i  (Proteintech)
96
Proteintech collagen type i
Collagen Type I, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 96 stars, based on 1 article reviews
collagen type i - by Bioz Stars, 2026-06
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syn1  (Proteintech)
95
Proteintech syn1
a , b , The representative immunoblots ( a ) and quantitative analyses ( b ) of Glun1, Glun2A, Glun2B and <t>Syn1</t> in the hippocampus of the WT, 3xTg-AD and 3xTg-AD+β-OHB mice, n = 4 per group. c , RT–qPCR assays mRNA expression of the Glun1, Glun2A, Glun2B and Syn1 in the hippocampus of the WT, 3xTg-AD and 3xTg-AD+β-OHB mice, n = 5 per group. d , ChIP–qPCR analysis of the enrichment of H3K9bhb at Glun1, Glun2A, Glun2B, Glun2C and Syn1 promoters in the hippocampus of the WT, 3xTg-AD and 3xTg-AD+β-OHB mice, n = 5 per group. e , f , Supplementing with β-OHB could increase the density of 3xTg-AD dendritic spines detected by Golgi-cox staining; the representative images ( e ) and quantitative analysis ( f ) of spine, n = 5 per group, three fields per mice. Scale bar, 5 μm. g – j , The Sholl analysis showed the synaptic complexity of neurons after supplementing with β-OHB in 3xTg-AD mice; the representative images ( g and i ) and the quantitative analysis ( h and j ), n = 5 per group, two fields per mice. Scale bar, 50 μm. Data are shown as mean ± s.e.m. One-way ANOVA followed by Bonferroni’s post hoc test for b – d and f . Two-way ANOVA followed by Bonferroni’s post hoc test for i and k . * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns, not significant.
Syn1, supplied by Proteintech, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 95 stars, based on 1 article reviews
syn1 - by Bioz Stars, 2026-06
95/100 stars
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rig  (Proteintech)
95
Proteintech rig
a , b , The representative immunoblots ( a ) and quantitative analyses ( b ) of Glun1, Glun2A, Glun2B and <t>Syn1</t> in the hippocampus of the WT, 3xTg-AD and 3xTg-AD+β-OHB mice, n = 4 per group. c , RT–qPCR assays mRNA expression of the Glun1, Glun2A, Glun2B and Syn1 in the hippocampus of the WT, 3xTg-AD and 3xTg-AD+β-OHB mice, n = 5 per group. d , ChIP–qPCR analysis of the enrichment of H3K9bhb at Glun1, Glun2A, Glun2B, Glun2C and Syn1 promoters in the hippocampus of the WT, 3xTg-AD and 3xTg-AD+β-OHB mice, n = 5 per group. e , f , Supplementing with β-OHB could increase the density of 3xTg-AD dendritic spines detected by Golgi-cox staining; the representative images ( e ) and quantitative analysis ( f ) of spine, n = 5 per group, three fields per mice. Scale bar, 5 μm. g – j , The Sholl analysis showed the synaptic complexity of neurons after supplementing with β-OHB in 3xTg-AD mice; the representative images ( g and i ) and the quantitative analysis ( h and j ), n = 5 per group, two fields per mice. Scale bar, 50 μm. Data are shown as mean ± s.e.m. One-way ANOVA followed by Bonferroni’s post hoc test for b – d and f . Two-way ANOVA followed by Bonferroni’s post hoc test for i and k . * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns, not significant.
Rig, supplied by Proteintech, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 95 stars, based on 1 article reviews
rig - by Bioz Stars, 2026-06
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anti tumor susceptibility gene 101  (Proteintech)
96
Proteintech anti tumor susceptibility gene 101
a , b , The representative immunoblots ( a ) and quantitative analyses ( b ) of Glun1, Glun2A, Glun2B and <t>Syn1</t> in the hippocampus of the WT, 3xTg-AD and 3xTg-AD+β-OHB mice, n = 4 per group. c , RT–qPCR assays mRNA expression of the Glun1, Glun2A, Glun2B and Syn1 in the hippocampus of the WT, 3xTg-AD and 3xTg-AD+β-OHB mice, n = 5 per group. d , ChIP–qPCR analysis of the enrichment of H3K9bhb at Glun1, Glun2A, Glun2B, Glun2C and Syn1 promoters in the hippocampus of the WT, 3xTg-AD and 3xTg-AD+β-OHB mice, n = 5 per group. e , f , Supplementing with β-OHB could increase the density of 3xTg-AD dendritic spines detected by Golgi-cox staining; the representative images ( e ) and quantitative analysis ( f ) of spine, n = 5 per group, three fields per mice. Scale bar, 5 μm. g – j , The Sholl analysis showed the synaptic complexity of neurons after supplementing with β-OHB in 3xTg-AD mice; the representative images ( g and i ) and the quantitative analysis ( h and j ), n = 5 per group, two fields per mice. Scale bar, 50 μm. Data are shown as mean ± s.e.m. One-way ANOVA followed by Bonferroni’s post hoc test for b – d and f . Two-way ANOVA followed by Bonferroni’s post hoc test for i and k . * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns, not significant.
Anti Tumor Susceptibility Gene 101, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 96 stars, based on 1 article reviews
anti tumor susceptibility gene 101 - by Bioz Stars, 2026-06
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annexin a1 monoclonal antibody  (Proteintech)
94
Proteintech annexin a1 monoclonal antibody
(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 <t>ANXA1</t> 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.
Annexin A1 Monoclonal Antibody, supplied by Proteintech, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 94 stars, based on 1 article reviews
annexin a1 monoclonal antibody - by Bioz Stars, 2026-06
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anti ikbα antibody  (Proteintech)
95
Proteintech anti ikbα antibody
miR-19-EXO dampened M1 macrophages polarization via inhibiting NF-κB canonical activation. (A) The procedure of THP1-based macrophage polarization assay. THP1 monocytes were treated with 100 ng/mL PMA for 48 h (for M0 induction), then treatment treated with 20 ng/mL LPS and 20 ng/mL IFN-γ for M1 subtype polarization, or 20 ng/mL IL4 and 20 ng/mL IL-13 or M2 subtype polarization, respectively. Representative morphology of M0, M1, M2 were recorded (scale bar = 100 μm). (B) and (D) RT-PCR analysis of M1 marker TNF-α , Il-6 and M2 marker Il-10 , CD163 in WT-EXO/miR-19-EXO treated M1 or M2 cells. (C) and (E) The macrophage polarization status in WT-EXO/miR-19-EXO treated M1/M2 cells was estimated by CD86 + and CD206 + cells ration using flow cytometry. (F) Thp1-M1 cells treated with miR-19-EXO or miR-19KD-EXO (100 μg/mL) for 24 h, then protein level of p47phox, NF-κB P65 and p-P65 were examined by immunoblotting. Thp1-M0 as control. (G) Thp1-M1 cell administrated with miR-19-EXO (100 μg/mL) for 24 h or NC were treated with CHX (100 μg/mL) for 0, 30, 60, 120, 240 min. Immunoblotting for <t>iKBα</t> expression levels in whole-cell extraction. (H) Thp1-M1 cell administrated with miR-19-EXO for 24 h or NC were treated with MG132 (10 mmol/L) for an extra 12h, then immunoblotting for iKBα protein levels in each group. (I) Thp1-M1 cell administrated with miR-19-EXO for 24 h or NC were immunoprecipitated <t>with</t> <t>IgG</t> control or iKBα antibody and then immunoblotted for Ubiquitin and iKBα. (J) Thp1-M1 cells treated with miR-19-EXO for 24 h were subjected to nuclear cytoplasmic separation assay, NF-κB P65 and IKKα level and translocation were examined by immunoblotting. LaminB1 was used as an internal control of nuclear protein, GAPDH as cytoplasmic protein control. (K) Schematic diagram for the mechanisms of IKK/NF-κB pathway activation suppressed by miR-19-EXO. (L)Thp1-M1 cells treated with miR-19-EXO or Thp1-M1 shp47phox cells or Thp1-M1 shp47phox cells treated with miR-19-EXO (100 μg/mL) for 24 h were detected the protein level of p47phox, NF-κB P65 and p-P65 by immunoblotting. Thp1-M1 as control (Ctrl). (M)Statistical differences were determined using unpaired Student's t -test between each 2 cohorts (ns = no significance, ∗p < 0.05, ∗∗p < 0.01).
Anti Ikbα Antibody, supplied by Proteintech, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 95 stars, based on 1 article reviews
anti ikbα antibody - by Bioz Stars, 2026-06
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sc 47681 anti mouse collectin proteintech cat  (Proteintech)
92
Proteintech sc 47681 anti mouse collectin proteintech cat
miR-19-EXO dampened M1 macrophages polarization via inhibiting NF-κB canonical activation. (A) The procedure of THP1-based macrophage polarization assay. THP1 monocytes were treated with 100 ng/mL PMA for 48 h (for M0 induction), then treatment treated with 20 ng/mL LPS and 20 ng/mL IFN-γ for M1 subtype polarization, or 20 ng/mL IL4 and 20 ng/mL IL-13 or M2 subtype polarization, respectively. Representative morphology of M0, M1, M2 were recorded (scale bar = 100 μm). (B) and (D) RT-PCR analysis of M1 marker TNF-α , Il-6 and M2 marker Il-10 , CD163 in WT-EXO/miR-19-EXO treated M1 or M2 cells. (C) and (E) The macrophage polarization status in WT-EXO/miR-19-EXO treated M1/M2 cells was estimated by CD86 + and CD206 + cells ration using flow cytometry. (F) Thp1-M1 cells treated with miR-19-EXO or miR-19KD-EXO (100 μg/mL) for 24 h, then protein level of p47phox, NF-κB P65 and p-P65 were examined by immunoblotting. Thp1-M0 as control. (G) Thp1-M1 cell administrated with miR-19-EXO (100 μg/mL) for 24 h or NC were treated with CHX (100 μg/mL) for 0, 30, 60, 120, 240 min. Immunoblotting for <t>iKBα</t> expression levels in whole-cell extraction. (H) Thp1-M1 cell administrated with miR-19-EXO for 24 h or NC were treated with MG132 (10 mmol/L) for an extra 12h, then immunoblotting for iKBα protein levels in each group. (I) Thp1-M1 cell administrated with miR-19-EXO for 24 h or NC were immunoprecipitated <t>with</t> <t>IgG</t> control or iKBα antibody and then immunoblotted for Ubiquitin and iKBα. (J) Thp1-M1 cells treated with miR-19-EXO for 24 h were subjected to nuclear cytoplasmic separation assay, NF-κB P65 and IKKα level and translocation were examined by immunoblotting. LaminB1 was used as an internal control of nuclear protein, GAPDH as cytoplasmic protein control. (K) Schematic diagram for the mechanisms of IKK/NF-κB pathway activation suppressed by miR-19-EXO. (L)Thp1-M1 cells treated with miR-19-EXO or Thp1-M1 shp47phox cells or Thp1-M1 shp47phox cells treated with miR-19-EXO (100 μg/mL) for 24 h were detected the protein level of p47phox, NF-κB P65 and p-P65 by immunoblotting. Thp1-M1 as control (Ctrl). (M)Statistical differences were determined using unpaired Student's t -test between each 2 cohorts (ns = no significance, ∗p < 0.05, ∗∗p < 0.01).
Sc 47681 Anti Mouse Collectin Proteintech Cat, supplied by Proteintech, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ndufb9  (Proteintech)
93
Proteintech ndufb9
miR-19-EXO dampened M1 macrophages polarization via inhibiting NF-κB canonical activation. (A) The procedure of THP1-based macrophage polarization assay. THP1 monocytes were treated with 100 ng/mL PMA for 48 h (for M0 induction), then treatment treated with 20 ng/mL LPS and 20 ng/mL IFN-γ for M1 subtype polarization, or 20 ng/mL IL4 and 20 ng/mL IL-13 or M2 subtype polarization, respectively. Representative morphology of M0, M1, M2 were recorded (scale bar = 100 μm). (B) and (D) RT-PCR analysis of M1 marker TNF-α , Il-6 and M2 marker Il-10 , CD163 in WT-EXO/miR-19-EXO treated M1 or M2 cells. (C) and (E) The macrophage polarization status in WT-EXO/miR-19-EXO treated M1/M2 cells was estimated by CD86 + and CD206 + cells ration using flow cytometry. (F) Thp1-M1 cells treated with miR-19-EXO or miR-19KD-EXO (100 μg/mL) for 24 h, then protein level of p47phox, NF-κB P65 and p-P65 were examined by immunoblotting. Thp1-M0 as control. (G) Thp1-M1 cell administrated with miR-19-EXO (100 μg/mL) for 24 h or NC were treated with CHX (100 μg/mL) for 0, 30, 60, 120, 240 min. Immunoblotting for <t>iKBα</t> expression levels in whole-cell extraction. (H) Thp1-M1 cell administrated with miR-19-EXO for 24 h or NC were treated with MG132 (10 mmol/L) for an extra 12h, then immunoblotting for iKBα protein levels in each group. (I) Thp1-M1 cell administrated with miR-19-EXO for 24 h or NC were immunoprecipitated <t>with</t> <t>IgG</t> control or iKBα antibody and then immunoblotted for Ubiquitin and iKBα. (J) Thp1-M1 cells treated with miR-19-EXO for 24 h were subjected to nuclear cytoplasmic separation assay, NF-κB P65 and IKKα level and translocation were examined by immunoblotting. LaminB1 was used as an internal control of nuclear protein, GAPDH as cytoplasmic protein control. (K) Schematic diagram for the mechanisms of IKK/NF-κB pathway activation suppressed by miR-19-EXO. (L)Thp1-M1 cells treated with miR-19-EXO or Thp1-M1 shp47phox cells or Thp1-M1 shp47phox cells treated with miR-19-EXO (100 μg/mL) for 24 h were detected the protein level of p47phox, NF-κB P65 and p-P65 by immunoblotting. Thp1-M1 as control (Ctrl). (M)Statistical differences were determined using unpaired Student's t -test between each 2 cohorts (ns = no significance, ∗p < 0.05, ∗∗p < 0.01).
Ndufb9, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ikkβ proteintech 15649 1 ap  (Proteintech)
94
Proteintech ikkβ proteintech 15649 1 ap
miR-19-EXO dampened M1 macrophages polarization via inhibiting NF-κB canonical activation. (A) The procedure of THP1-based macrophage polarization assay. THP1 monocytes were treated with 100 ng/mL PMA for 48 h (for M0 induction), then treatment treated with 20 ng/mL LPS and 20 ng/mL IFN-γ for M1 subtype polarization, or 20 ng/mL IL4 and 20 ng/mL IL-13 or M2 subtype polarization, respectively. Representative morphology of M0, M1, M2 were recorded (scale bar = 100 μm). (B) and (D) RT-PCR analysis of M1 marker TNF-α , Il-6 and M2 marker Il-10 , CD163 in WT-EXO/miR-19-EXO treated M1 or M2 cells. (C) and (E) The macrophage polarization status in WT-EXO/miR-19-EXO treated M1/M2 cells was estimated by CD86 + and CD206 + cells ration using flow cytometry. (F) Thp1-M1 cells treated with miR-19-EXO or miR-19KD-EXO (100 μg/mL) for 24 h, then protein level of p47phox, NF-κB P65 and p-P65 were examined by immunoblotting. Thp1-M0 as control. (G) Thp1-M1 cell administrated with miR-19-EXO (100 μg/mL) for 24 h or NC were treated with CHX (100 μg/mL) for 0, 30, 60, 120, 240 min. Immunoblotting for <t>iKBα</t> expression levels in whole-cell extraction. (H) Thp1-M1 cell administrated with miR-19-EXO for 24 h or NC were treated with MG132 (10 mmol/L) for an extra 12h, then immunoblotting for iKBα protein levels in each group. (I) Thp1-M1 cell administrated with miR-19-EXO for 24 h or NC were immunoprecipitated <t>with</t> <t>IgG</t> control or iKBα antibody and then immunoblotted for Ubiquitin and iKBα. (J) Thp1-M1 cells treated with miR-19-EXO for 24 h were subjected to nuclear cytoplasmic separation assay, NF-κB P65 and IKKα level and translocation were examined by immunoblotting. LaminB1 was used as an internal control of nuclear protein, GAPDH as cytoplasmic protein control. (K) Schematic diagram for the mechanisms of IKK/NF-κB pathway activation suppressed by miR-19-EXO. (L)Thp1-M1 cells treated with miR-19-EXO or Thp1-M1 shp47phox cells or Thp1-M1 shp47phox cells treated with miR-19-EXO (100 μg/mL) for 24 h were detected the protein level of p47phox, NF-κB P65 and p-P65 by immunoblotting. Thp1-M1 as control (Ctrl). (M)Statistical differences were determined using unpaired Student's t -test between each 2 cohorts (ns = no significance, ∗p < 0.05, ∗∗p < 0.01).
Ikkβ Proteintech 15649 1 Ap, supplied by Proteintech, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ikkβ proteintech 15649 1 ap - by Bioz Stars, 2026-06
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rabbit cpt1b antibody wb proteintech 22170 1 ap  (Proteintech)
96
Proteintech rabbit cpt1b antibody wb proteintech 22170 1 ap
miR-19-EXO dampened M1 macrophages polarization via inhibiting NF-κB canonical activation. (A) The procedure of THP1-based macrophage polarization assay. THP1 monocytes were treated with 100 ng/mL PMA for 48 h (for M0 induction), then treatment treated with 20 ng/mL LPS and 20 ng/mL IFN-γ for M1 subtype polarization, or 20 ng/mL IL4 and 20 ng/mL IL-13 or M2 subtype polarization, respectively. Representative morphology of M0, M1, M2 were recorded (scale bar = 100 μm). (B) and (D) RT-PCR analysis of M1 marker TNF-α , Il-6 and M2 marker Il-10 , CD163 in WT-EXO/miR-19-EXO treated M1 or M2 cells. (C) and (E) The macrophage polarization status in WT-EXO/miR-19-EXO treated M1/M2 cells was estimated by CD86 + and CD206 + cells ration using flow cytometry. (F) Thp1-M1 cells treated with miR-19-EXO or miR-19KD-EXO (100 μg/mL) for 24 h, then protein level of p47phox, NF-κB P65 and p-P65 were examined by immunoblotting. Thp1-M0 as control. (G) Thp1-M1 cell administrated with miR-19-EXO (100 μg/mL) for 24 h or NC were treated with CHX (100 μg/mL) for 0, 30, 60, 120, 240 min. Immunoblotting for <t>iKBα</t> expression levels in whole-cell extraction. (H) Thp1-M1 cell administrated with miR-19-EXO for 24 h or NC were treated with MG132 (10 mmol/L) for an extra 12h, then immunoblotting for iKBα protein levels in each group. (I) Thp1-M1 cell administrated with miR-19-EXO for 24 h or NC were immunoprecipitated <t>with</t> <t>IgG</t> control or iKBα antibody and then immunoblotted for Ubiquitin and iKBα. (J) Thp1-M1 cells treated with miR-19-EXO for 24 h were subjected to nuclear cytoplasmic separation assay, NF-κB P65 and IKKα level and translocation were examined by immunoblotting. LaminB1 was used as an internal control of nuclear protein, GAPDH as cytoplasmic protein control. (K) Schematic diagram for the mechanisms of IKK/NF-κB pathway activation suppressed by miR-19-EXO. (L)Thp1-M1 cells treated with miR-19-EXO or Thp1-M1 shp47phox cells or Thp1-M1 shp47phox cells treated with miR-19-EXO (100 μg/mL) for 24 h were detected the protein level of p47phox, NF-κB P65 and p-P65 by immunoblotting. Thp1-M1 as control (Ctrl). (M)Statistical differences were determined using unpaired Student's t -test between each 2 cohorts (ns = no significance, ∗p < 0.05, ∗∗p < 0.01).
Rabbit Cpt1b Antibody Wb Proteintech 22170 1 Ap, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rabbit cpt1b antibody wb proteintech 22170 1 ap - by Bioz Stars, 2026-06
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germany 27124 1 ap igf1r  (Proteintech)
94
Proteintech germany 27124 1 ap igf1r
miR-19-EXO dampened M1 macrophages polarization via inhibiting NF-κB canonical activation. (A) The procedure of THP1-based macrophage polarization assay. THP1 monocytes were treated with 100 ng/mL PMA for 48 h (for M0 induction), then treatment treated with 20 ng/mL LPS and 20 ng/mL IFN-γ for M1 subtype polarization, or 20 ng/mL IL4 and 20 ng/mL IL-13 or M2 subtype polarization, respectively. Representative morphology of M0, M1, M2 were recorded (scale bar = 100 μm). (B) and (D) RT-PCR analysis of M1 marker TNF-α , Il-6 and M2 marker Il-10 , CD163 in WT-EXO/miR-19-EXO treated M1 or M2 cells. (C) and (E) The macrophage polarization status in WT-EXO/miR-19-EXO treated M1/M2 cells was estimated by CD86 + and CD206 + cells ration using flow cytometry. (F) Thp1-M1 cells treated with miR-19-EXO or miR-19KD-EXO (100 μg/mL) for 24 h, then protein level of p47phox, NF-κB P65 and p-P65 were examined by immunoblotting. Thp1-M0 as control. (G) Thp1-M1 cell administrated with miR-19-EXO (100 μg/mL) for 24 h or NC were treated with CHX (100 μg/mL) for 0, 30, 60, 120, 240 min. Immunoblotting for <t>iKBα</t> expression levels in whole-cell extraction. (H) Thp1-M1 cell administrated with miR-19-EXO for 24 h or NC were treated with MG132 (10 mmol/L) for an extra 12h, then immunoblotting for iKBα protein levels in each group. (I) Thp1-M1 cell administrated with miR-19-EXO for 24 h or NC were immunoprecipitated <t>with</t> <t>IgG</t> control or iKBα antibody and then immunoblotted for Ubiquitin and iKBα. (J) Thp1-M1 cells treated with miR-19-EXO for 24 h were subjected to nuclear cytoplasmic separation assay, NF-κB P65 and IKKα level and translocation were examined by immunoblotting. LaminB1 was used as an internal control of nuclear protein, GAPDH as cytoplasmic protein control. (K) Schematic diagram for the mechanisms of IKK/NF-κB pathway activation suppressed by miR-19-EXO. (L)Thp1-M1 cells treated with miR-19-EXO or Thp1-M1 shp47phox cells or Thp1-M1 shp47phox cells treated with miR-19-EXO (100 μg/mL) for 24 h were detected the protein level of p47phox, NF-κB P65 and p-P65 by immunoblotting. Thp1-M1 as control (Ctrl). (M)Statistical differences were determined using unpaired Student's t -test between each 2 cohorts (ns = no significance, ∗p < 0.05, ∗∗p < 0.01).
Germany 27124 1 Ap Igf1r, supplied by Proteintech, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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a , b , The representative immunoblots ( a ) and quantitative analyses ( b ) of Glun1, Glun2A, Glun2B and Syn1 in the hippocampus of the WT, 3xTg-AD and 3xTg-AD+β-OHB mice, n = 4 per group. c , RT–qPCR assays mRNA expression of the Glun1, Glun2A, Glun2B and Syn1 in the hippocampus of the WT, 3xTg-AD and 3xTg-AD+β-OHB mice, n = 5 per group. d , ChIP–qPCR analysis of the enrichment of H3K9bhb at Glun1, Glun2A, Glun2B, Glun2C and Syn1 promoters in the hippocampus of the WT, 3xTg-AD and 3xTg-AD+β-OHB mice, n = 5 per group. e , f , Supplementing with β-OHB could increase the density of 3xTg-AD dendritic spines detected by Golgi-cox staining; the representative images ( e ) and quantitative analysis ( f ) of spine, n = 5 per group, three fields per mice. Scale bar, 5 μm. g – j , The Sholl analysis showed the synaptic complexity of neurons after supplementing with β-OHB in 3xTg-AD mice; the representative images ( g and i ) and the quantitative analysis ( h and j ), n = 5 per group, two fields per mice. Scale bar, 50 μm. Data are shown as mean ± s.e.m. One-way ANOVA followed by Bonferroni’s post hoc test for b – d and f . Two-way ANOVA followed by Bonferroni’s post hoc test for i and k . * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns, not significant.

Journal: Experimental & Molecular Medicine

Article Title: HMGCS2-dependent β-OHB/H3K9bhb ameliorates synaptic plasticity and cognition in Alzheimer’s disease

doi: 10.1038/s12276-026-01664-9

Figure Lengend Snippet: a , b , The representative immunoblots ( a ) and quantitative analyses ( b ) of Glun1, Glun2A, Glun2B and Syn1 in the hippocampus of the WT, 3xTg-AD and 3xTg-AD+β-OHB mice, n = 4 per group. c , RT–qPCR assays mRNA expression of the Glun1, Glun2A, Glun2B and Syn1 in the hippocampus of the WT, 3xTg-AD and 3xTg-AD+β-OHB mice, n = 5 per group. d , ChIP–qPCR analysis of the enrichment of H3K9bhb at Glun1, Glun2A, Glun2B, Glun2C and Syn1 promoters in the hippocampus of the WT, 3xTg-AD and 3xTg-AD+β-OHB mice, n = 5 per group. e , f , Supplementing with β-OHB could increase the density of 3xTg-AD dendritic spines detected by Golgi-cox staining; the representative images ( e ) and quantitative analysis ( f ) of spine, n = 5 per group, three fields per mice. Scale bar, 5 μm. g – j , The Sholl analysis showed the synaptic complexity of neurons after supplementing with β-OHB in 3xTg-AD mice; the representative images ( g and i ) and the quantitative analysis ( h and j ), n = 5 per group, two fields per mice. Scale bar, 50 μm. Data are shown as mean ± s.e.m. One-way ANOVA followed by Bonferroni’s post hoc test for b – d and f . Two-way ANOVA followed by Bonferroni’s post hoc test for i and k . * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns, not significant.

Article Snippet: SYN1 , Synapsin-1 , Poly- , 1:1000 , Proteintech , 20258-1-AP.

Techniques: Western Blot, Quantitative RT-PCR, Expressing, ChIP-qPCR, Staining

a – c , The HMGCS2 upregulation promotes the protein ( a and b ) and mRNA ( c ) expression of H3K9bhb, Glun1, Glun2A, Glun2B, Syn1 and PSD95, n = 4 or 5 per group. d , e , ChIP–qPCR analyses of the enrichment of H3K9bhb at Glun1, Glun2A, Glun2B and Syn1 promoters in the primary neurons of the WT, 3xTg-AD and 3xTg-AD + HMGCS2 mice n = 5 per group ( d ) and representative gel images from ChIP–qPCR assays ( e ). f – j , The HMGCS2 upregulation promotes the expression of Syn1 (scale bar, 25 μm) ( f ); n = 10 cells per group in MAP2 immunofluorescence ( g ) and quantitative analysis ( h ), n = 10 cells per group and SYP ( i and j ), n = 10 cells per group, scale bar, 15 μm. Data are shown as mean ± s.e.m. One-way ANOVA followed by Bonferroni’s post hoc test for b – d and j . Two-way ANOVA followed by Bonferroni’s post hoc test for h . * P < 0.05, ** P < 0.01 , *** P < 0.001, **** P < 0.0001; ns, not significant.

Journal: Experimental & Molecular Medicine

Article Title: HMGCS2-dependent β-OHB/H3K9bhb ameliorates synaptic plasticity and cognition in Alzheimer’s disease

doi: 10.1038/s12276-026-01664-9

Figure Lengend Snippet: a – c , The HMGCS2 upregulation promotes the protein ( a and b ) and mRNA ( c ) expression of H3K9bhb, Glun1, Glun2A, Glun2B, Syn1 and PSD95, n = 4 or 5 per group. d , e , ChIP–qPCR analyses of the enrichment of H3K9bhb at Glun1, Glun2A, Glun2B and Syn1 promoters in the primary neurons of the WT, 3xTg-AD and 3xTg-AD + HMGCS2 mice n = 5 per group ( d ) and representative gel images from ChIP–qPCR assays ( e ). f – j , The HMGCS2 upregulation promotes the expression of Syn1 (scale bar, 25 μm) ( f ); n = 10 cells per group in MAP2 immunofluorescence ( g ) and quantitative analysis ( h ), n = 10 cells per group and SYP ( i and j ), n = 10 cells per group, scale bar, 15 μm. Data are shown as mean ± s.e.m. One-way ANOVA followed by Bonferroni’s post hoc test for b – d and j . Two-way ANOVA followed by Bonferroni’s post hoc test for h . * P < 0.05, ** P < 0.01 , *** P < 0.001, **** P < 0.0001; ns, not significant.

Article Snippet: SYN1 , Synapsin-1 , Poly- , 1:1000 , Proteintech , 20258-1-AP.

Techniques: Expressing, ChIP-qPCR, Immunofluorescence

a , b , A western blot analysis ( a ) of hippocampal lysates shows that HMGCS2 upregulation increases the protein levels ( b ) of H3K9bhb, Glun1, Glun2A, Glun2B, Syn1 and PSD95, n = 3 per group. c , The ChIP–qPCR analysis of H3K9bhb enrichment at the promoters of Glun2A , Glun2B , Syn1 and PSD95 in the four groups, n = 5 per group. d , The mRNA levels of Glun1, Glun2A, Glun2B, Syn1 and PSD95 in the hippocampus, as determined by RT–qPCR, n = 5 per group. e , f , Golgi staining reveals increased dendritic spine density in 3xTg-AD mice following overexpression of HMGCS2; representative images ( e ) and quantification ( f ) are shown, n = 5 per group, three fields per mice. Scale bar, 5 μm. g – j , A behavioral assessment of spatial learning and memory using the MWM, NOR and contextual fear conditioning tests: area under the curve (AUC) of escape latency during MWM training of day 1–6 ( g ), escape latency on day 7 of the MWM test ( h ), NOR discrimination index ( i ), freezing time on day 7 in the contextual fear conditioning test ( j ), n = 8 per group. Data are shown as mean ± s.e.m. One-way ANOVA followed by Bonferroni’s post hoc test for b – d , f and g – j . * P < 0.05, ** P < 0.01 , *** P < 0.001, **** P < 0.0001; ns, not significant.

Journal: Experimental & Molecular Medicine

Article Title: HMGCS2-dependent β-OHB/H3K9bhb ameliorates synaptic plasticity and cognition in Alzheimer’s disease

doi: 10.1038/s12276-026-01664-9

Figure Lengend Snippet: a , b , A western blot analysis ( a ) of hippocampal lysates shows that HMGCS2 upregulation increases the protein levels ( b ) of H3K9bhb, Glun1, Glun2A, Glun2B, Syn1 and PSD95, n = 3 per group. c , The ChIP–qPCR analysis of H3K9bhb enrichment at the promoters of Glun2A , Glun2B , Syn1 and PSD95 in the four groups, n = 5 per group. d , The mRNA levels of Glun1, Glun2A, Glun2B, Syn1 and PSD95 in the hippocampus, as determined by RT–qPCR, n = 5 per group. e , f , Golgi staining reveals increased dendritic spine density in 3xTg-AD mice following overexpression of HMGCS2; representative images ( e ) and quantification ( f ) are shown, n = 5 per group, three fields per mice. Scale bar, 5 μm. g – j , A behavioral assessment of spatial learning and memory using the MWM, NOR and contextual fear conditioning tests: area under the curve (AUC) of escape latency during MWM training of day 1–6 ( g ), escape latency on day 7 of the MWM test ( h ), NOR discrimination index ( i ), freezing time on day 7 in the contextual fear conditioning test ( j ), n = 8 per group. Data are shown as mean ± s.e.m. One-way ANOVA followed by Bonferroni’s post hoc test for b – d , f and g – j . * P < 0.05, ** P < 0.01 , *** P < 0.001, **** P < 0.0001; ns, not significant.

Article Snippet: SYN1 , Synapsin-1 , Poly- , 1:1000 , Proteintech , 20258-1-AP.

Techniques: Western Blot, ChIP-qPCR, Quantitative RT-PCR, Staining, Over Expression

(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.

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

(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.

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

(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.

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

(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.

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

(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.

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

(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.

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

(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.

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

(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.

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

miR-19-EXO dampened M1 macrophages polarization via inhibiting NF-κB canonical activation. (A) The procedure of THP1-based macrophage polarization assay. THP1 monocytes were treated with 100 ng/mL PMA for 48 h (for M0 induction), then treatment treated with 20 ng/mL LPS and 20 ng/mL IFN-γ for M1 subtype polarization, or 20 ng/mL IL4 and 20 ng/mL IL-13 or M2 subtype polarization, respectively. Representative morphology of M0, M1, M2 were recorded (scale bar = 100 μm). (B) and (D) RT-PCR analysis of M1 marker TNF-α , Il-6 and M2 marker Il-10 , CD163 in WT-EXO/miR-19-EXO treated M1 or M2 cells. (C) and (E) The macrophage polarization status in WT-EXO/miR-19-EXO treated M1/M2 cells was estimated by CD86 + and CD206 + cells ration using flow cytometry. (F) Thp1-M1 cells treated with miR-19-EXO or miR-19KD-EXO (100 μg/mL) for 24 h, then protein level of p47phox, NF-κB P65 and p-P65 were examined by immunoblotting. Thp1-M0 as control. (G) Thp1-M1 cell administrated with miR-19-EXO (100 μg/mL) for 24 h or NC were treated with CHX (100 μg/mL) for 0, 30, 60, 120, 240 min. Immunoblotting for iKBα expression levels in whole-cell extraction. (H) Thp1-M1 cell administrated with miR-19-EXO for 24 h or NC were treated with MG132 (10 mmol/L) for an extra 12h, then immunoblotting for iKBα protein levels in each group. (I) Thp1-M1 cell administrated with miR-19-EXO for 24 h or NC were immunoprecipitated with IgG control or iKBα antibody and then immunoblotted for Ubiquitin and iKBα. (J) Thp1-M1 cells treated with miR-19-EXO for 24 h were subjected to nuclear cytoplasmic separation assay, NF-κB P65 and IKKα level and translocation were examined by immunoblotting. LaminB1 was used as an internal control of nuclear protein, GAPDH as cytoplasmic protein control. (K) Schematic diagram for the mechanisms of IKK/NF-κB pathway activation suppressed by miR-19-EXO. (L)Thp1-M1 cells treated with miR-19-EXO or Thp1-M1 shp47phox cells or Thp1-M1 shp47phox cells treated with miR-19-EXO (100 μg/mL) for 24 h were detected the protein level of p47phox, NF-κB P65 and p-P65 by immunoblotting. Thp1-M1 as control (Ctrl). (M)Statistical differences were determined using unpaired Student's t -test between each 2 cohorts (ns = no significance, ∗p < 0.05, ∗∗p < 0.01).

Journal: Materials Today Bio

Article Title: miR-19b-3p engineered adipose-derived stem cell exosomes attenuate acute liver failure via promoting tissue repair and microenvironment remodeling

doi: 10.1016/j.mtbio.2025.102697

Figure Lengend Snippet: miR-19-EXO dampened M1 macrophages polarization via inhibiting NF-κB canonical activation. (A) The procedure of THP1-based macrophage polarization assay. THP1 monocytes were treated with 100 ng/mL PMA for 48 h (for M0 induction), then treatment treated with 20 ng/mL LPS and 20 ng/mL IFN-γ for M1 subtype polarization, or 20 ng/mL IL4 and 20 ng/mL IL-13 or M2 subtype polarization, respectively. Representative morphology of M0, M1, M2 were recorded (scale bar = 100 μm). (B) and (D) RT-PCR analysis of M1 marker TNF-α , Il-6 and M2 marker Il-10 , CD163 in WT-EXO/miR-19-EXO treated M1 or M2 cells. (C) and (E) The macrophage polarization status in WT-EXO/miR-19-EXO treated M1/M2 cells was estimated by CD86 + and CD206 + cells ration using flow cytometry. (F) Thp1-M1 cells treated with miR-19-EXO or miR-19KD-EXO (100 μg/mL) for 24 h, then protein level of p47phox, NF-κB P65 and p-P65 were examined by immunoblotting. Thp1-M0 as control. (G) Thp1-M1 cell administrated with miR-19-EXO (100 μg/mL) for 24 h or NC were treated with CHX (100 μg/mL) for 0, 30, 60, 120, 240 min. Immunoblotting for iKBα expression levels in whole-cell extraction. (H) Thp1-M1 cell administrated with miR-19-EXO for 24 h or NC were treated with MG132 (10 mmol/L) for an extra 12h, then immunoblotting for iKBα protein levels in each group. (I) Thp1-M1 cell administrated with miR-19-EXO for 24 h or NC were immunoprecipitated with IgG control or iKBα antibody and then immunoblotted for Ubiquitin and iKBα. (J) Thp1-M1 cells treated with miR-19-EXO for 24 h were subjected to nuclear cytoplasmic separation assay, NF-κB P65 and IKKα level and translocation were examined by immunoblotting. LaminB1 was used as an internal control of nuclear protein, GAPDH as cytoplasmic protein control. (K) Schematic diagram for the mechanisms of IKK/NF-κB pathway activation suppressed by miR-19-EXO. (L)Thp1-M1 cells treated with miR-19-EXO or Thp1-M1 shp47phox cells or Thp1-M1 shp47phox cells treated with miR-19-EXO (100 μg/mL) for 24 h were detected the protein level of p47phox, NF-κB P65 and p-P65 by immunoblotting. Thp1-M1 as control (Ctrl). (M)Statistical differences were determined using unpaired Student's t -test between each 2 cohorts (ns = no significance, ∗p < 0.05, ∗∗p < 0.01).

Article Snippet: Next, the protein A/G agarose beads were pre-binded with 500 μL of anti-iKBα antibody (1:250, Proteintech, China) or rabbit IgG antibody (1:250, CST, USA) by rotating 1h at room temperature, then washed the antibody connected-beads for 3 times.

Techniques: Activation Assay, Reverse Transcription Polymerase Chain Reaction, Marker, Flow Cytometry, Western Blot, Control, Expressing, Extraction, Immunoprecipitation, Ubiquitin Proteomics, Translocation Assay