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Thermo Fisher apc-cy7 anti-cd8
Apc Cy7 Anti Cd8, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/apc-cy7+anti-cd8/anti+cd4+fitc/pmc12171361-107-4-25
Average 90 stars, based on 1 article reviews

apc-cy7 anti-cd8 - by Bioz Stars, 2026-11

90/100 stars

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Article Title: Dual variants of uncertain significance in a case of hyper-IgM syndrome: implications for diagnosis and management
Article Snippet: Cells were stained with APC-Cy7 anti-CD8, then fixed with BD Cytofix/Cytoperm (BD Bioscience) and stained with AF488 anti-CD4, PE anti-IFNγ, BV650 anti-TNFα, and APC anti-IL-2 (eBioscience).

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Figure 4. StCl-GI_GII impairs tumor growth by reactivating the B16F10 melanoma tumor immune environment. a) Timeline (days) of tumor inoculation and immunization (s.c. = subcutaneous). b) Representative images of tumors collected on day twenty-one after post-tumor inoculation. c) Tumor growth curve. Data presented as the mean ± SEM (7< N < 10). Two-way ANOVA. P values correspond to tumor volume at day twenty-one after tumor inoculation. d–g) Tumor-inﬁltrating immune cell populations for d) CD3+ T cells, e) CD3+PD-1+ T cells, f) CTLs, and g) CD3+CD8+PD-1+ T cells. Tumors isolated on day twenty-one after tumor inoculation, and quantiﬁcation performed by ﬂow cytometry. Data presented as mean ± SD, n ≥3 animals. h) Functional assessment of antigen-speciﬁc T cells shown by a tetramer staining assay. Percent of gp100-speciﬁc CD8+ T cells in the spleen. Data presented as the mean ± SD, n = 3. Statistics: one-way analysis of variance (ANOVA) followed by Tukey post-hoc test; i) Heatmap of diﬀerentially-expressed genes in tumors between PBS-treated and StCl-GI_GII-treated mice. Yellow and blue represent a high and low expression. The graded color represents the Z-score to compare the number of reads by row. More details in Tables S3–S5 (Supporting Information). j) Biodistribution of StCI-GI_GII-Cy5 in the lymph nodes: i) Low magniﬁcation image showing the architecture of an inguinal lymph node and StCI-GI_GII-Cy5 biodistribution. ii) Magniﬁcation of i) around the aﬀerent and eﬀerent lymphatic vessels. iii) and v) Higher magniﬁcation of ii showing colocalization of StCl-GI_GII-Cy5 and lymphatic endothelial cells (LECs) (Yellow arrows). iv,v) Higher magniﬁcation of ii showing colocalization between StCl-GI_GII-Cy5 and blood endothelial cells (BECs) (white arrows). The dashed line shows BECs (in red) colocalizing with StCl-GI_GII-Cy5 in a more exposed image.

Journal: Advanced materials (Deerfield Beach, Fla.)

Article Title: Off-The-Shelf Multivalent Nanoconjugate Cancer Vaccine Rescues Host Immune Response against Melanoma.

doi: 10.1002/adma.202417348

Figure Lengend Snippet: Figure 4. StCl-GI_GII impairs tumor growth by reactivating the B16F10 melanoma tumor immune environment. a) Timeline (days) of tumor inoculation and immunization (s.c. = subcutaneous). b) Representative images of tumors collected on day twenty-one after post-tumor inoculation. c) Tumor growth curve. Data presented as the mean ± SEM (7< N < 10). Two-way ANOVA. P values correspond to tumor volume at day twenty-one after tumor inoculation. d–g) Tumor-inﬁltrating immune cell populations for d) CD3+ T cells, e) CD3+PD-1+ T cells, f) CTLs, and g) CD3+CD8+PD-1+ T cells. Tumors isolated on day twenty-one after tumor inoculation, and quantiﬁcation performed by ﬂow cytometry. Data presented as mean ± SD, n ≥3 animals. h) Functional assessment of antigen-speciﬁc T cells shown by a tetramer staining assay. Percent of gp100-speciﬁc CD8+ T cells in the spleen. Data presented as the mean ± SD, n = 3. Statistics: one-way analysis of variance (ANOVA) followed by Tukey post-hoc test; i) Heatmap of diﬀerentially-expressed genes in tumors between PBS-treated and StCl-GI_GII-treated mice. Yellow and blue represent a high and low expression. The graded color represents the Z-score to compare the number of reads by row. More details in Tables S3–S5 (Supporting Information). j) Biodistribution of StCI-GI_GII-Cy5 in the lymph nodes: i) Low magniﬁcation image showing the architecture of an inguinal lymph node and StCI-GI_GII-Cy5 biodistribution. ii) Magniﬁcation of i) around the aﬀerent and eﬀerent lymphatic vessels. iii) and v) Higher magniﬁcation of ii showing colocalization of StCl-GI_GII-Cy5 and lymphatic endothelial cells (LECs) (Yellow arrows). iv,v) Higher magniﬁcation of ii showing colocalization between StCl-GI_GII-Cy5 and blood endothelial cells (BECs) (white arrows). The dashed line shows BECs (in red) colocalizing with StCl-GI_GII-Cy5 in a more exposed image.

Article Snippet: Panel: T Lymphocytes / Tregs/ Cytokines CD8± CD3-APC-Cy7, Miltenyi [Cat.# 130-119-793, mouse, clone: REA641]; CD8aFITC, Biolegend [Cat.# 100 706, mouse, clone: 53–6.7]; CD4-PerCPCy5.5, Biolegend [Cat.# 100 434, mouse, clone: GK1.5]; CD25-PE-Cy7, Miltenyi [Cat.# 130-123-893 (30 μg in 200 μL), mouse, clone: REA568]; CD279 (PD1)-PE, Miltenyi [Cat.# 130-111-953, mouse, clone: REA802]; CD107b-Alexa647, Biolegend, Cat.# 108 512, mouse, clone: M3/84]; CD127-PE-Cy5, Biolegend, Cat.# 135 015, mouse, clone: A7R34]; IL-2BV71, BioLegend [Cat.# 503 837, mouse, clone: JES6-5H4]; Anti-IFN-γBV785, Biolegend [Cat# 505 838, mouse, clone: XMG1.2]; Anti-TNF-αBV421, Biolegend, Cat.# 506 328, mouse, clone: MP6-XT22] Panel: Dendritic cell (DC) / Myeloid-derived suppressor cells (MDSC) / Tumor-associated macrophages (TAM) CD11c-PE-Cy5, Biolegend [Cat.# 117 316, mouse, clone: N418]; CD11bAPC-Cy7, Miltenyi [Cat.# 130-113-803, mouse, clone: REA592]; Anti-MHC II-VioBlue, Miltenyi [Cat.# 130-112-394, mouse, clone: REA813];CD8a-FITC, Biolegend [Cat.# 100 706, mouse, clone: 53–6.7]; CD103-PerCP-Cy5.5), Biolegend [Cat.# 121 416, mouse, clone: 2E7]; Anti-XCR1-PE, Miltenyi [Cat.# 130-111-372 mouse, clone: REA707]; Anti-Gr1-APC, Miltenyi [Cat.# 130-112-307, mouse, clone: REA810]; Anti-F4/80-PE-Cy7, Miltenyi [Cat.# 130-118-459 mouse, clone: REA126]; CD206 (MMR)-BV711, BioLegend [Cat.# 141 727, mouse, clone: C068C2] Panel: Lymphatic endothelial cells (LECs) / Blood vascular endothelial cells (BECs) CD45-PerCP-Cy5.5, Miltenyi [Cat.# 130-110-801, mouse, clone: REA737]; CD31-PE, Biolegend, Cat.# 102 408, mouse, clone: 390]; Podoplanin (PDPN, gp38)-APC-Cy7, BioLegend [Cat.# 127 417, mouse, clone: 8.1.1]; CD273 (PD-L2)-PE-Cy7, Miltenyi [Cat.# 130-102-875, mouse, clone: MIH37]; CD274 (PD-L1)-BV711, BioLegend [Cat.# 12 431 922] Panel: Human T Lymphocytes CD45-APC-Vio 770, Miltenyi [Cat.# 130-110-635, human, clone: REA747]; CD3- PerCP-Cy5.5, Invitrogen [Cat.# 45-0037-42, human; CD8-VioBlue, Miltenyi [Cat.# 130-110-683, human, clone: REA734]; CD4-FITC, Miltenyi [Cat.# 130-114-585, human, clone: REA623]; CD279 (PD1)-PE, Miltenyi [Cat.# 130-120-382, human, clone: REA1165].

Techniques: Isolation, Cytometry, Functional Assay, Staining, Expressing

Figure 5. Anti-tumor therapeutic synergy between a multivalent nanoconjugate cancer vaccine and 𝛼-PD-1. a) Timeline (days) of tumor inoculation and immunization (s.c. = subcutaneous). b) Representative images of tumors collected on day twenty-one after post-tumor inoculation. c) Individual tumor volumes at day twenty-one after tumor inoculation. Data presented as mean ± SEM (10 < N < 13). d) Tumor growth curve. Data presented as mean ± SEM (10 < N < 13). One-way ANOVA. e) Kaplan-Meier overall survival for mice inoculated with B16F10 melanoma cells. Data presented as mean ± SEM (7 < N <10); f–p) Tumor-inﬁltrating immune cell populations for f) CD3+, g) CD8+, h) ratio of CD8+ T: T reg cells, i) TAMs, j) PD-1 expressing CD8+ cells, k) Tregs, l) DCs, (m and o) BECs, and (n and p) LECs. Tumors isolated on day twenty-one after tumor inoculation, and quantiﬁcation performed by ﬂow cytometry. Data presented as the mean ± SD, n = 3. Statistics: one-way analysis of variance (ANOVA) followed by Tukey post-hoc test.; q) Quantiﬁcation of Casp3 and r) Ki67after immunostaining at the experimental endpoint. Data presented as mean ± SEM, n ≥3 animals.

Journal: Advanced materials (Deerfield Beach, Fla.)

Article Title: Off-The-Shelf Multivalent Nanoconjugate Cancer Vaccine Rescues Host Immune Response against Melanoma.

doi: 10.1002/adma.202417348

Figure Lengend Snippet: Figure 5. Anti-tumor therapeutic synergy between a multivalent nanoconjugate cancer vaccine and 𝛼-PD-1. a) Timeline (days) of tumor inoculation and immunization (s.c. = subcutaneous). b) Representative images of tumors collected on day twenty-one after post-tumor inoculation. c) Individual tumor volumes at day twenty-one after tumor inoculation. Data presented as mean ± SEM (10 < N < 13). d) Tumor growth curve. Data presented as mean ± SEM (10 < N < 13). One-way ANOVA. e) Kaplan-Meier overall survival for mice inoculated with B16F10 melanoma cells. Data presented as mean ± SEM (7 < N <10); f–p) Tumor-inﬁltrating immune cell populations for f) CD3+, g) CD8+, h) ratio of CD8+ T: T reg cells, i) TAMs, j) PD-1 expressing CD8+ cells, k) Tregs, l) DCs, (m and o) BECs, and (n and p) LECs. Tumors isolated on day twenty-one after tumor inoculation, and quantiﬁcation performed by ﬂow cytometry. Data presented as the mean ± SD, n = 3. Statistics: one-way analysis of variance (ANOVA) followed by Tukey post-hoc test.; q) Quantiﬁcation of Casp3 and r) Ki67after immunostaining at the experimental endpoint. Data presented as mean ± SEM, n ≥3 animals.

Techniques: Expressing, Isolation, Cytometry, Immunostaining

Figure 6. Validating the immune-mediated anti-tumor eﬀect of the multivalent nanoconjugate cancer vaccine platform in a melanoma PDX mouse model. a) Representative images of gp100 expression in a murine tumor from a PDX tumor derived from a melanoma patient. Images acquired at 8x magniﬁcation. Scale bar 400 μm b) Timeline (days) of tumor inoculation, immunization, and 𝛼PD-1 treatment (s.c. = subcutaneous). c) Tumor growth curve. Data presented as mean ± SEM (6 < N < 7). One-way ANOVA. P values correspond to tumor volume on days ﬁfteen/seventeen after treatment initiation. Representative images of xenograft tumors at day seventeen post-treatment initiation. d–h) Percentage of (d) CD3+ T cells within the tumor and (e) CD8+, (f) CD4+, (g) CD3+PD-1+, and (h) CD8+PD-1+ T cells within the spleen at day seventeen post-treatment initiation. Data presented as the mean ± SD, n = 3. Statistics: one-way analysis of variance (ANOVA) followed by Tukey post-hoc test; i–k) Quantiﬁcation of tumor-inﬁltrating (i) CD3+, (j) CD4+, and (k) CD8+ T-cell populations, in addition to l) Caspase-3 and m) Ki67 by immunostaining at day seventeen. Data presented as mean ± SEM, n ≥3 animals. n) Representative images of tumor immunostaining for CD3, CD4, CD8, Caspase-3, and Ki67 in all treated mice. Scale bar 200 μm o) Representative images of H&E tumor staining from mice treated with PBS, StCl-GI_GII, and StCl-GI_GII + ɑPD-1. Extended tumor necrosis in the three groups (i, ii, and iii), with active apoptosis, inﬂammatory intra, and peri-tumor response in StCl-GI_GII and StCl-GI_GII + ɑPD1 groups (ii, iii). HE,10X-20X magniﬁcation; scale bar 120–250 μm.

Journal: Advanced materials (Deerfield Beach, Fla.)

Article Title: Off-The-Shelf Multivalent Nanoconjugate Cancer Vaccine Rescues Host Immune Response against Melanoma.

doi: 10.1002/adma.202417348

Figure Lengend Snippet: Figure 6. Validating the immune-mediated anti-tumor eﬀect of the multivalent nanoconjugate cancer vaccine platform in a melanoma PDX mouse model. a) Representative images of gp100 expression in a murine tumor from a PDX tumor derived from a melanoma patient. Images acquired at 8x magniﬁcation. Scale bar 400 μm b) Timeline (days) of tumor inoculation, immunization, and 𝛼PD-1 treatment (s.c. = subcutaneous). c) Tumor growth curve. Data presented as mean ± SEM (6 < N < 7). One-way ANOVA. P values correspond to tumor volume on days ﬁfteen/seventeen after treatment initiation. Representative images of xenograft tumors at day seventeen post-treatment initiation. d–h) Percentage of (d) CD3+ T cells within the tumor and (e) CD8+, (f) CD4+, (g) CD3+PD-1+, and (h) CD8+PD-1+ T cells within the spleen at day seventeen post-treatment initiation. Data presented as the mean ± SD, n = 3. Statistics: one-way analysis of variance (ANOVA) followed by Tukey post-hoc test; i–k) Quantiﬁcation of tumor-inﬁltrating (i) CD3+, (j) CD4+, and (k) CD8+ T-cell populations, in addition to l) Caspase-3 and m) Ki67 by immunostaining at day seventeen. Data presented as mean ± SEM, n ≥3 animals. n) Representative images of tumor immunostaining for CD3, CD4, CD8, Caspase-3, and Ki67 in all treated mice. Scale bar 200 μm o) Representative images of H&E tumor staining from mice treated with PBS, StCl-GI_GII, and StCl-GI_GII + ɑPD-1. Extended tumor necrosis in the three groups (i, ii, and iii), with active apoptosis, inﬂammatory intra, and peri-tumor response in StCl-GI_GII and StCl-GI_GII + ɑPD1 groups (ii, iii). HE,10X-20X magniﬁcation; scale bar 120–250 μm.

Techniques: Expressing, Derivative Assay, Immunostaining, Staining

(a) Graphs showing percentage of positive mouse and human CD45 cells, percentage of human CD3, CD8, CD19 and human CD45 cell counts in weekly in peripheral blood (tail vein bleeds) for the two PBMC donor groups (PBMC Donor A, top; PBMC Donor B, bottom). Error bars represent mean±SEM of 4-8 mice per group.

Journal: bioRxiv

Article Title: Immune and Mutational Profile of Gene-Edited Low-Immunogenic Human Primary Cholangiocyte Organoids

doi: 10.1101/2025.01.20.628680

Figure Lengend Snippet: (a) Graphs showing percentage of positive mouse and human CD45 cells, percentage of human CD3, CD8, CD19 and human CD45 cell counts in weekly in peripheral blood (tail vein bleeds) for the two PBMC donor groups (PBMC Donor A, top; PBMC Donor B, bottom). Error bars represent mean±SEM of 4-8 mice per group.

Article Snippet: Humanization was evaluated weekly by flow cytometry of tail-vein bleeds collected in heparin-coated tubes (Sarstedt Ltd, 20.1309), followed by red cell lysis removal (StemCell Technologies, 07850) according to manufacturer’s instructions and using the following conjugated antibodies: mouse anti-human CD45-FITC (1:50, Thermo Fisher Scientific, 11-0459-42, clone [HI30]), mouse anti-mouse CD45.1-PE-Cy7 (1:100, Thermo Fisher Scientific, 25-0453-82, clone [A20]), mouse anti-human CD3-APC (1:50, Biolegend, clone [UCHT1]), mouse anti-human CD19-PE (1:50, Biolegend, 363004, clone [SJ25C1]), and mouse anti-human CD8-APC/Cy7 (1:50, Thermo Fisher Scientific, A15441, clone [RFT8]).

Techniques:

Lethally-irradiated B6 mice were grafted with a 1:1 mixture of B6 (H-2 b ) and B6D2F1 (H-2 bd ) bone marrow and co-injected with WT or HP1α-deficient naive CD4 + T cells alone or in the presence of ex vivo expanded WT Treg. To control engraftment, a group of mice was also injected solely with the bone marrow cell mixture. a Experimental model. b Representative dot-plots showing the frequency of syngeneic (H-2K b+ ) and semi-allogeneic (H-2K bd+ ) cells in the blood 21 days after engraftment. c Percentage of semi-allogeneic (H-2K d+ ) cells among peripheral blood mononuclear cells (PBMC) 21 days after engraftment. Data show mean ± SEM of 3 to 6 independent experiments. Each symbol represents the mean of an experiment. P values were calculated using multiple unpaired t-tests with Holm-Šidák correction. Source data are provided in the Source data file.

Journal: Nature Communications

Article Title: Heterochromatic gene silencing controls CD4 + T cell susceptibility to regulatory T cell-mediated suppression in a murine allograft model

doi: 10.1038/s41467-025-55848-4

Figure Lengend Snippet: Lethally-irradiated B6 mice were grafted with a 1:1 mixture of B6 (H-2 b ) and B6D2F1 (H-2 bd ) bone marrow and co-injected with WT or HP1α-deficient naive CD4 + T cells alone or in the presence of ex vivo expanded WT Treg. To control engraftment, a group of mice was also injected solely with the bone marrow cell mixture. a Experimental model. b Representative dot-plots showing the frequency of syngeneic (H-2K b+ ) and semi-allogeneic (H-2K bd+ ) cells in the blood 21 days after engraftment. c Percentage of semi-allogeneic (H-2K d+ ) cells among peripheral blood mononuclear cells (PBMC) 21 days after engraftment. Data show mean ± SEM of 3 to 6 independent experiments. Each symbol represents the mean of an experiment. P values were calculated using multiple unpaired t-tests with Holm-Šidák correction. Source data are provided in the Source data file.

Article Snippet: Extracellular staining was performed with BV421-coupled anti-TCRβ (H57-597, BD Biosciences), BUV395-labeled anti-CD4, APC-Cy7-conjugated anti-CD8 (eBioH35-17.2, Thermo Fisher Scientific), PE-Cy7-conjugated anti-CD44 (IM7, BD Biosciences), APC-coupled anti-CD62L (MEL-14, Thermo Fisher Scientific) and PE-conjugated anti-CD25 antibodies.

Techniques: Irradiation, Injection, Ex Vivo, Control

Lethally-irradiated B6 mice were grafted with a 1:1 mixture of B6 and B6D2F1 bone marrow and co-injected with WT or HP1α-deficient naive CD4 + T cells alone or in the presence of ex vivo expanded WT Treg. a Representative dot-plots showing 21 days after engraftment the percentage of WT and HP1α KO TCR Vβ6 + CD4 + T cells producing IFN-γ and IL-17A. b – d Percentage of WT or HP1α KO TCR Vβ6 + CD4 + T cells producing IFN-γ (b), IL-17A ( c ) or both ( d ) 21 days after engraftment. e Volcano plot showing results of differential gene expression analyses between Treg that had been co-injected with WT or HP1α KO naive CD4 + T cells. Red and black dots represent genes with higher expression in Treg co-injected with HP1α KO or WT cells, respectively. Gray dots represent genes that failed to reach the FDR threshold of 0.05 and the absolute log2 fold change threshold of 1. f Absolute number of spleen cells 21 days after engraftment. Data are mean ± SD of three (Tconv) or seven (Tconv + Treg) biological replicates from two independent experiments. g Percentage of Tconv, defined as CD4 + CD45.1 - CD45.2 + H-2K d- Thy1.1 - , in the spleen 21 days after engraftment. Data are mean ± SEM of three independent experiments. h Percentage of allospecific Tconv, defined as CD4 + CD45.1 - CD45.2 + H-2K d- Thy1.1 - TCRVβ6 + , in the spleen 21 days after engraftment. Data are mean ± SEM of three independent experiments. i, j Percentage of CD73 + FR4 + ( i ) or Foxp3 + ( j ) cells among WT or HP1α KO TCR Vβ6 + CD4 + T cells, as determined in the spleen 21 days after engraftment. b – d and g – j Data show mean ± SEM of 3 independent experiments. Each symbol represents the mean of an experiment. P values were calculated using multiple unpaired t test with Holm-Šidák correction. Source data are provided in the Source data file.

Journal: Nature Communications

Article Title: Heterochromatic gene silencing controls CD4 + T cell susceptibility to regulatory T cell-mediated suppression in a murine allograft model

doi: 10.1038/s41467-025-55848-4

Figure Lengend Snippet: Lethally-irradiated B6 mice were grafted with a 1:1 mixture of B6 and B6D2F1 bone marrow and co-injected with WT or HP1α-deficient naive CD4 + T cells alone or in the presence of ex vivo expanded WT Treg. a Representative dot-plots showing 21 days after engraftment the percentage of WT and HP1α KO TCR Vβ6 + CD4 + T cells producing IFN-γ and IL-17A. b – d Percentage of WT or HP1α KO TCR Vβ6 + CD4 + T cells producing IFN-γ (b), IL-17A ( c ) or both ( d ) 21 days after engraftment. e Volcano plot showing results of differential gene expression analyses between Treg that had been co-injected with WT or HP1α KO naive CD4 + T cells. Red and black dots represent genes with higher expression in Treg co-injected with HP1α KO or WT cells, respectively. Gray dots represent genes that failed to reach the FDR threshold of 0.05 and the absolute log2 fold change threshold of 1. f Absolute number of spleen cells 21 days after engraftment. Data are mean ± SD of three (Tconv) or seven (Tconv + Treg) biological replicates from two independent experiments. g Percentage of Tconv, defined as CD4 + CD45.1 - CD45.2 + H-2K d- Thy1.1 - , in the spleen 21 days after engraftment. Data are mean ± SEM of three independent experiments. h Percentage of allospecific Tconv, defined as CD4 + CD45.1 - CD45.2 + H-2K d- Thy1.1 - TCRVβ6 + , in the spleen 21 days after engraftment. Data are mean ± SEM of three independent experiments. i, j Percentage of CD73 + FR4 + ( i ) or Foxp3 + ( j ) cells among WT or HP1α KO TCR Vβ6 + CD4 + T cells, as determined in the spleen 21 days after engraftment. b – d and g – j Data show mean ± SEM of 3 independent experiments. Each symbol represents the mean of an experiment. P values were calculated using multiple unpaired t test with Holm-Šidák correction. Source data are provided in the Source data file.

Techniques: Irradiation, Injection, Ex Vivo, Expressing

a Differentially-expressed genes (DEG) between WT and HP1α-deficient T cells. b GSEA of KEGG pathways performed using transcriptomes of HP1α-deficient and WT Tconv exposed to Treg. c GO enrichment analyses of genes more highly expressed in WT Tconv than in HP1α KO Tconv exposed to Treg. d GSEA of exhaustion signature genes performed using transcriptomes of HP1α-deficient and WT Tconv exposed to Treg. e Number of differentially-accessible peaks (DAP) between WT and HP1α-deficient T cells. f Relationship between the genes more highly expressed and the genes associated with peaks more open in HP1α KO Tconv than in WT Tconv exposed to Treg. g Expression levels of the 54 genes selected from the intersection in f . Horizontal bars represent the median, and the top and bottom of the boxes the upper and lower quartiles, respectively. The whiskers go from the minimum to the lower quartile and from the upper quartile to the maximum. Statistical significance was calculated using the Pairwise Wilcoxon Rank Sum Test (two-tailed). h GO enrichment analyses of the 54 genes selected from the intersection in f . i Th1 signature enrichment analyses of the 54 genes selected from the intersection in f . j ATAC–seq and RNA-seq tracks at the Il2 locus. k UMAP plot of tumor-infiltrating CD4 + T lymphocytes from melanoma patients . Cells are colored based on 6 clusters defined using the SNN algorithm. l Relative proportions of CD4 + T cell subsets in patients who responded (R) or not (NR) to immunotherapy. m Teff:Tex ratio in R and NR patients. Statistical significance was calculated using Wilcoxon rank sum Test (two-tailed). Horizontal bars represent the median, and the top and bottom of the boxes the upper and lower quartiles, respectively. The whiskers go from the minimum to the lower quartile and from the upper quartile to the maximum. n GSEA of genes downregulated in HP1α-deficient vs control Tconv performed using transcriptomes of human Teff and Tex. Significance was estimated using rank-based gene permutations. Source data are provided in the Source data file.

Journal: Nature Communications

Article Title: Heterochromatic gene silencing controls CD4 + T cell susceptibility to regulatory T cell-mediated suppression in a murine allograft model

doi: 10.1038/s41467-025-55848-4

Figure Lengend Snippet: a Differentially-expressed genes (DEG) between WT and HP1α-deficient T cells. b GSEA of KEGG pathways performed using transcriptomes of HP1α-deficient and WT Tconv exposed to Treg. c GO enrichment analyses of genes more highly expressed in WT Tconv than in HP1α KO Tconv exposed to Treg. d GSEA of exhaustion signature genes performed using transcriptomes of HP1α-deficient and WT Tconv exposed to Treg. e Number of differentially-accessible peaks (DAP) between WT and HP1α-deficient T cells. f Relationship between the genes more highly expressed and the genes associated with peaks more open in HP1α KO Tconv than in WT Tconv exposed to Treg. g Expression levels of the 54 genes selected from the intersection in f . Horizontal bars represent the median, and the top and bottom of the boxes the upper and lower quartiles, respectively. The whiskers go from the minimum to the lower quartile and from the upper quartile to the maximum. Statistical significance was calculated using the Pairwise Wilcoxon Rank Sum Test (two-tailed). h GO enrichment analyses of the 54 genes selected from the intersection in f . i Th1 signature enrichment analyses of the 54 genes selected from the intersection in f . j ATAC–seq and RNA-seq tracks at the Il2 locus. k UMAP plot of tumor-infiltrating CD4 + T lymphocytes from melanoma patients . Cells are colored based on 6 clusters defined using the SNN algorithm. l Relative proportions of CD4 + T cell subsets in patients who responded (R) or not (NR) to immunotherapy. m Teff:Tex ratio in R and NR patients. Statistical significance was calculated using Wilcoxon rank sum Test (two-tailed). Horizontal bars represent the median, and the top and bottom of the boxes the upper and lower quartiles, respectively. The whiskers go from the minimum to the lower quartile and from the upper quartile to the maximum. n GSEA of genes downregulated in HP1α-deficient vs control Tconv performed using transcriptomes of human Teff and Tex. Significance was estimated using rank-based gene permutations. Source data are provided in the Source data file.

Techniques: Expressing, Two Tailed Test, RNA Sequencing Assay, Control

Lethally-irradiated B6 mice were grafted with a 1:1 mixture of B6 and B6D2F1 bone marrow and co-injected with WT or HP1α KO ( a, b ) or SUV39H1 KO ( c, d ) naive CD4 + T cells alone or in the presence of ex vivo expanded WT Treg. To control engraftment, a group of mice was also injected solely with the bone marrow cell mixture. a, c Representative dot-plots showing the frequency of syngeneic (H-2K b+ ) and semi-allogeneic (H-2K bd+ ) cells in the blood 21 days after engraftment. b, d Percentage of semi-allogeneic (H-2K d+ ) cells among PBMC 21 days after engraftment. Data show mean ± SEM of 3 to 8 independent experiments. Each symbol represents the mean of an experiment. P values were calculated using multiple unpaired t test with Holm-Šidák correction. Source data are provided in the Source data file.

Journal: Nature Communications

Article Title: Heterochromatic gene silencing controls CD4 + T cell susceptibility to regulatory T cell-mediated suppression in a murine allograft model

doi: 10.1038/s41467-025-55848-4

Figure Lengend Snippet: Lethally-irradiated B6 mice were grafted with a 1:1 mixture of B6 and B6D2F1 bone marrow and co-injected with WT or HP1α KO ( a, b ) or SUV39H1 KO ( c, d ) naive CD4 + T cells alone or in the presence of ex vivo expanded WT Treg. To control engraftment, a group of mice was also injected solely with the bone marrow cell mixture. a, c Representative dot-plots showing the frequency of syngeneic (H-2K b+ ) and semi-allogeneic (H-2K bd+ ) cells in the blood 21 days after engraftment. b, d Percentage of semi-allogeneic (H-2K d+ ) cells among PBMC 21 days after engraftment. Data show mean ± SEM of 3 to 8 independent experiments. Each symbol represents the mean of an experiment. P values were calculated using multiple unpaired t test with Holm-Šidák correction. Source data are provided in the Source data file.

Techniques: Irradiation, Injection, Ex Vivo, Control

Lethally-irradiated B6 mice were grafted with a 1:1 mixture of B6 and B6D2F1 bone marrow and co-injected with WT (a-d and I, j) or HP1γ-deficient ( a–j ) naive CD4 + T cells alone or in the presence of ex vivo expanded WT Treg. a Representative dot-plots showing 21 days after engraftment the percentage of WT and HP1γ KO TCR Vβ6 + Tconv producing IFN-γ and IL-17A. b, c Percentage of WT or HP1γ KO TCR Vβ6 + Tconv producing IFN-γ ( b ) or IL-17A ( c ) 21 days after engraftment. d Percentage of CD73 + FR4 + cells among WT or HP1γ KO TCR Vβ6 + Tconv, as determined in the spleen 21 days after engraftment. e Percentage of cytokine-producing cells among HP1γ KO TCR Vβ6 + Tconv 21 days post-engraftment. The analysis was performed on total Tconv from mice injected with Tconv only, or on anergic (A) or non-anergic (N-A) Tconv from mice injected with Tconv and Treg. ( f ) Representative dot-plots showing 21 days after engraftment the percentage of HP1γ KO TCR Vβ6 + Tconv expressing PD-1 and TIGIT. g, h Percentage of PD-1 + , TIGIT + , LAG-3 + ( g ) or PD-1 + TIGIT + LAG-3 + ( h ) cells among HP1γ KO TCR Vβ6 + Tconv 21 days after engraftment. i Percentage of Foxp3 + cells among WT or HP1γ KO TCR Vβ6 + Tconv, as determined in the spleen 21 days after engraftment. j Percentage of spleen Foxp3 + cells among anergic or non-anergic WT and HP1γ KO TCR Vβ6 + Tconv 21 days after engraftment. b – d and I, j Data are represented as mean ± SEM of three independent experiments. Each symbol represents the mean of an experiment. P values were calculated using multiple unpaired t test with Holm-Šidák correction. e, g, h Floating bar charts represent the mean of the biological replicates as well as the minimum and maximum values. Each symbol represents individual biological replicates. P values were calculated using unpaired t test (two-tailed). Source data are provided in the Source data file.

Journal: Nature Communications

Article Title: Heterochromatic gene silencing controls CD4 + T cell susceptibility to regulatory T cell-mediated suppression in a murine allograft model

doi: 10.1038/s41467-025-55848-4

Figure Lengend Snippet: Lethally-irradiated B6 mice were grafted with a 1:1 mixture of B6 and B6D2F1 bone marrow and co-injected with WT (a-d and I, j) or HP1γ-deficient ( a–j ) naive CD4 + T cells alone or in the presence of ex vivo expanded WT Treg. a Representative dot-plots showing 21 days after engraftment the percentage of WT and HP1γ KO TCR Vβ6 + Tconv producing IFN-γ and IL-17A. b, c Percentage of WT or HP1γ KO TCR Vβ6 + Tconv producing IFN-γ ( b ) or IL-17A ( c ) 21 days after engraftment. d Percentage of CD73 + FR4 + cells among WT or HP1γ KO TCR Vβ6 + Tconv, as determined in the spleen 21 days after engraftment. e Percentage of cytokine-producing cells among HP1γ KO TCR Vβ6 + Tconv 21 days post-engraftment. The analysis was performed on total Tconv from mice injected with Tconv only, or on anergic (A) or non-anergic (N-A) Tconv from mice injected with Tconv and Treg. ( f ) Representative dot-plots showing 21 days after engraftment the percentage of HP1γ KO TCR Vβ6 + Tconv expressing PD-1 and TIGIT. g, h Percentage of PD-1 + , TIGIT + , LAG-3 + ( g ) or PD-1 + TIGIT + LAG-3 + ( h ) cells among HP1γ KO TCR Vβ6 + Tconv 21 days after engraftment. i Percentage of Foxp3 + cells among WT or HP1γ KO TCR Vβ6 + Tconv, as determined in the spleen 21 days after engraftment. j Percentage of spleen Foxp3 + cells among anergic or non-anergic WT and HP1γ KO TCR Vβ6 + Tconv 21 days after engraftment. b – d and I, j Data are represented as mean ± SEM of three independent experiments. Each symbol represents the mean of an experiment. P values were calculated using multiple unpaired t test with Holm-Šidák correction. e, g, h Floating bar charts represent the mean of the biological replicates as well as the minimum and maximum values. Each symbol represents individual biological replicates. P values were calculated using unpaired t test (two-tailed). Source data are provided in the Source data file.

Techniques: Irradiation, Injection, Ex Vivo, Expressing, Two Tailed Test

a Pattern of expression of genes overexpressed in Treg-exposed HP1γ KO Tconv compared with WT Tconv. ( b ) Cluster 2 genes expression in indicated Tconv populations. Horizontal bars represent the median, and the top and bottom of the box the upper and lower quartile, respectively. The whiskers go from the minimum to the lower quartile and from the upper quartile to the maximum. Statistical significance was calculated using the Pairwise Wilcoxon Rank Sum Test (two-tailed). c Exhaustion signature enrichment analyses of cluster 2 genes. d GO enrichment analyses of cluster 2 genes. e Euler diagram showing the relationship between cluster 2 genes and those associated with peaks more open in Treg-exposed HP1γ KO Tconv than WT Tconv. Exhaustion signature enrichment analysis was run on genes common to both groups. f ATAC–seq and RNA-seq tracks at the Lag3 locus. g ATAC-seq signal at peaks associated with cluster 2 genes. Peaks were divided into two sets according to whether or not they were differentially open in WT and KO T cells exposed to Treg. h–k PD-1 and LAG3 expression on naive CD4 + T cells before or after two days of culture with anti-CD3 antibody. h, j Representative histograms showing the expression of PD-1 ( h ) or LAG3 ( j ) in naive and activated T cells. i, k Percentage of activated T cells expressing PD-1 ( i ) or LAG3 ( k ) (left) and average immune checkpoint expression per cell (right). l Representative dot-plots showing PD-1 versus LAG3 expression by ex vivo stimulated CD4 + T cells with or without (control) DMSO or chaetocin. m, n Percentage of CD4 + T cells, either ex vivo activated and treated or not (control) with DMSO or chaetocin, expressing PD-1 ( m ) or LAG-3 ( n ). I, k, m, n Data show mean ± SD of biological replicates from 4 independent experiments. Each symbol represents an individual biological replicate. P values were calculated using unpaired t test (two-tailed). Source data are provided in the Source data file.

Journal: Nature Communications

Article Title: Heterochromatic gene silencing controls CD4 + T cell susceptibility to regulatory T cell-mediated suppression in a murine allograft model

doi: 10.1038/s41467-025-55848-4

Figure Lengend Snippet: a Pattern of expression of genes overexpressed in Treg-exposed HP1γ KO Tconv compared with WT Tconv. ( b ) Cluster 2 genes expression in indicated Tconv populations. Horizontal bars represent the median, and the top and bottom of the box the upper and lower quartile, respectively. The whiskers go from the minimum to the lower quartile and from the upper quartile to the maximum. Statistical significance was calculated using the Pairwise Wilcoxon Rank Sum Test (two-tailed). c Exhaustion signature enrichment analyses of cluster 2 genes. d GO enrichment analyses of cluster 2 genes. e Euler diagram showing the relationship between cluster 2 genes and those associated with peaks more open in Treg-exposed HP1γ KO Tconv than WT Tconv. Exhaustion signature enrichment analysis was run on genes common to both groups. f ATAC–seq and RNA-seq tracks at the Lag3 locus. g ATAC-seq signal at peaks associated with cluster 2 genes. Peaks were divided into two sets according to whether or not they were differentially open in WT and KO T cells exposed to Treg. h–k PD-1 and LAG3 expression on naive CD4 + T cells before or after two days of culture with anti-CD3 antibody. h, j Representative histograms showing the expression of PD-1 ( h ) or LAG3 ( j ) in naive and activated T cells. i, k Percentage of activated T cells expressing PD-1 ( i ) or LAG3 ( k ) (left) and average immune checkpoint expression per cell (right). l Representative dot-plots showing PD-1 versus LAG3 expression by ex vivo stimulated CD4 + T cells with or without (control) DMSO or chaetocin. m, n Percentage of CD4 + T cells, either ex vivo activated and treated or not (control) with DMSO or chaetocin, expressing PD-1 ( m ) or LAG-3 ( n ). I, k, m, n Data show mean ± SD of biological replicates from 4 independent experiments. Each symbol represents an individual biological replicate. P values were calculated using unpaired t test (two-tailed). Source data are provided in the Source data file.

Techniques: Expressing, Two Tailed Test, RNA Sequencing Assay, Ex Vivo, Control

HLA-B7 - WT or HP1γ-deficient human naive CD4 + T cells were injected i.v . into sublethally-irradiated NSG mice with or without ex vivo expanded HLA-B7 + human Treg. Three weeks after injection, splenocytes were isolated and the xenogeneic T cell response was analyzed by flow cytometry. a Experimental model. b , c Following genome editing by CRISPR-Cas9, the expression level of HP1γ was determined in WT and HP1γ KO human CD4 + T cells. A representative western-blot ( b ) and normalized expression levels for each experiment ( c ) are shown. d Representative dot-plots showing the percentage of HLA-B7 - WT and HP1γ KO human Tconv producing IFN-γ and GM-CSF. e – g Percentage of HLA-B7 - WT and HP1γ KO human Tconv producing IFN-γ ( e ), GM-CSF ( f ) or Granzyme B ( g ). h Expression level of T-bet in HLA-B7 - WT and HP1γ KO human CD4 + T cells. i , j Percentage of HLA-B7 - WT and HP1γ KO human CD4 + T cells producing IL-10 (i) or expressing Foxp3 ( j ). Data show values for biological replicates from two ( e – j ) or three ( c ) independent experiments. Horizontal bars represent mean ± SD ( e – j ) or mean ± SEM ( c ). P values were calculated using two-tailed unpaired t tests. Source data are provided in the Source data file.

Journal: Nature Communications

Article Title: Heterochromatic gene silencing controls CD4 + T cell susceptibility to regulatory T cell-mediated suppression in a murine allograft model

doi: 10.1038/s41467-025-55848-4

Figure Lengend Snippet: HLA-B7 - WT or HP1γ-deficient human naive CD4 + T cells were injected i.v . into sublethally-irradiated NSG mice with or without ex vivo expanded HLA-B7 + human Treg. Three weeks after injection, splenocytes were isolated and the xenogeneic T cell response was analyzed by flow cytometry. a Experimental model. b , c Following genome editing by CRISPR-Cas9, the expression level of HP1γ was determined in WT and HP1γ KO human CD4 + T cells. A representative western-blot ( b ) and normalized expression levels for each experiment ( c ) are shown. d Representative dot-plots showing the percentage of HLA-B7 - WT and HP1γ KO human Tconv producing IFN-γ and GM-CSF. e – g Percentage of HLA-B7 - WT and HP1γ KO human Tconv producing IFN-γ ( e ), GM-CSF ( f ) or Granzyme B ( g ). h Expression level of T-bet in HLA-B7 - WT and HP1γ KO human CD4 + T cells. i , j Percentage of HLA-B7 - WT and HP1γ KO human CD4 + T cells producing IL-10 (i) or expressing Foxp3 ( j ). Data show values for biological replicates from two ( e – j ) or three ( c ) independent experiments. Horizontal bars represent mean ± SD ( e – j ) or mean ± SEM ( c ). P values were calculated using two-tailed unpaired t tests. Source data are provided in the Source data file.

Techniques: Injection, Irradiation, Ex Vivo, Isolation, Flow Cytometry, CRISPR, Expressing, Western Blot, Two Tailed Test

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