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a , b MC38 tumor-bearing male mice were treated i.p. with CTR, 40 μg anti-mouse CTLA4 antibody (mCTLA-4), or equimolar mouse ProCTLA-4 (mProCTLA-4) ( n = 5/group) on day 11 after tumor inoculation. 2 days later, Treg cells from the tumor, spleen, and dLN were analyzed by flow cytometry. The representative plot ( a ) and the frequency of Treg cells ( b ) from different treatment groups were shown. c – g Tumor-infiltrating CD45 + <t>immune</t> cells isolated from MC38 tumor-bearing male mice with different treatment (CTR, mCTLA-4, mProCTLA-4) were analyzed by scRNA-seq. c UMAP visualization of merged total CD45 + cells from three treatment groups. d Bar plot depicting the <t>cell</t> percentages of each cell type as a proportion of total cells. Feature plot illustrating gene expression of ICOS ( e ) and CTLA-4 ( f ) in Foxp3 + Treg cells. g Bar plot depicting the cell percentages of ICOS high Treg cells within total CD3 + T cells. h – j MC38 tumor-bearing male mice were treated i.p. with CTR, 10 μg mCTLA4, or equimolar mProCTLA4 on day 11 after tumor inoculation. 2 days after the treatment, the total Treg cells and ICOS high Treg cells ( h ) from tumors were analyzed by flow cytometry. The frequency and absolute number of total Treg cells ( i , j ) and ICOS high Treg cells ( k , l ) was quantified. m Female Rag1 -/- mice ( n = 5/group) were i.v. transferred with Tregs. 3 days post-transfer, the mice were challenged s.c. with MC38-OVA. Activated OT-Ι T cells were transferred 4 days later. 5 days after OT-Ι transfer, mice were treated with 100 μg mProCTLA-4 every three days for three times, and the tumor curve was monitored. <t>Data</t> in panels ( b , i – m ) representative of two independent experiments. Statistical analysis was performed using ordinary one-way ANOVA with multiple comparisons test ( b, i – l ), or two-way ANOVA with Tukey’s multiple comparisons test ( m ).
Murine Immune Cell Reference Data, supplied by Celldex Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Alithea Genomics bulk rna seq data analysis alithea genomics performed organoid lysis
a , b MC38 tumor-bearing male mice were treated i.p. with CTR, 40 μg anti-mouse CTLA4 antibody (mCTLA-4), or equimolar mouse ProCTLA-4 (mProCTLA-4) ( n = 5/group) on day 11 after tumor inoculation. 2 days later, Treg cells from the tumor, spleen, and dLN were analyzed by flow cytometry. The representative plot ( a ) and the frequency of Treg cells ( b ) from different treatment groups were shown. c – g Tumor-infiltrating CD45 + <t>immune</t> cells isolated from MC38 tumor-bearing male mice with different treatment (CTR, mCTLA-4, mProCTLA-4) were analyzed by scRNA-seq. c UMAP visualization of merged total CD45 + cells from three treatment groups. d Bar plot depicting the <t>cell</t> percentages of each cell type as a proportion of total cells. Feature plot illustrating gene expression of ICOS ( e ) and CTLA-4 ( f ) in Foxp3 + Treg cells. g Bar plot depicting the cell percentages of ICOS high Treg cells within total CD3 + T cells. h – j MC38 tumor-bearing male mice were treated i.p. with CTR, 10 μg mCTLA4, or equimolar mProCTLA4 on day 11 after tumor inoculation. 2 days after the treatment, the total Treg cells and ICOS high Treg cells ( h ) from tumors were analyzed by flow cytometry. The frequency and absolute number of total Treg cells ( i , j ) and ICOS high Treg cells ( k , l ) was quantified. m Female Rag1 -/- mice ( n = 5/group) were i.v. transferred with Tregs. 3 days post-transfer, the mice were challenged s.c. with MC38-OVA. Activated OT-Ι T cells were transferred 4 days later. 5 days after OT-Ι transfer, mice were treated with 100 μg mProCTLA-4 every three days for three times, and the tumor curve was monitored. <t>Data</t> in panels ( b , i – m ) representative of two independent experiments. Statistical analysis was performed using ordinary one-way ANOVA with multiple comparisons test ( b, i – l ), or two-way ANOVA with Tukey’s multiple comparisons test ( m ).
Bulk Rna Seq Data Analysis Alithea Genomics Performed Organoid Lysis, supplied by Alithea Genomics, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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a , b MC38 tumor-bearing male mice were treated i.p. with CTR, 40 μg anti-mouse CTLA4 antibody (mCTLA-4), or equimolar mouse ProCTLA-4 (mProCTLA-4) ( n = 5/group) on day 11 after tumor inoculation. 2 days later, Treg cells from the tumor, spleen, and dLN were analyzed by flow cytometry. The representative plot ( a ) and the frequency of Treg cells ( b ) from different treatment groups were shown. c – g Tumor-infiltrating CD45 + immune cells isolated from MC38 tumor-bearing male mice with different treatment (CTR, mCTLA-4, mProCTLA-4) were analyzed by scRNA-seq. c UMAP visualization of merged total CD45 + cells from three treatment groups. d Bar plot depicting the cell percentages of each cell type as a proportion of total cells. Feature plot illustrating gene expression of ICOS ( e ) and CTLA-4 ( f ) in Foxp3 + Treg cells. g Bar plot depicting the cell percentages of ICOS high Treg cells within total CD3 + T cells. h – j MC38 tumor-bearing male mice were treated i.p. with CTR, 10 μg mCTLA4, or equimolar mProCTLA4 on day 11 after tumor inoculation. 2 days after the treatment, the total Treg cells and ICOS high Treg cells ( h ) from tumors were analyzed by flow cytometry. The frequency and absolute number of total Treg cells ( i , j ) and ICOS high Treg cells ( k , l ) was quantified. m Female Rag1 -/- mice ( n = 5/group) were i.v. transferred with Tregs. 3 days post-transfer, the mice were challenged s.c. with MC38-OVA. Activated OT-Ι T cells were transferred 4 days later. 5 days after OT-Ι transfer, mice were treated with 100 μg mProCTLA-4 every three days for three times, and the tumor curve was monitored. Data in panels ( b , i – m ) representative of two independent experiments. Statistical analysis was performed using ordinary one-way ANOVA with multiple comparisons test ( b, i – l ), or two-way ANOVA with Tukey’s multiple comparisons test ( m ).

Journal: Nature Communications

Article Title: A next-generation anti-CTLA-4 probody mitigates toxicity and enhances anti-tumor immunity in mice

doi: 10.1038/s41467-025-64081-y

Figure Lengend Snippet: a , b MC38 tumor-bearing male mice were treated i.p. with CTR, 40 μg anti-mouse CTLA4 antibody (mCTLA-4), or equimolar mouse ProCTLA-4 (mProCTLA-4) ( n = 5/group) on day 11 after tumor inoculation. 2 days later, Treg cells from the tumor, spleen, and dLN were analyzed by flow cytometry. The representative plot ( a ) and the frequency of Treg cells ( b ) from different treatment groups were shown. c – g Tumor-infiltrating CD45 + immune cells isolated from MC38 tumor-bearing male mice with different treatment (CTR, mCTLA-4, mProCTLA-4) were analyzed by scRNA-seq. c UMAP visualization of merged total CD45 + cells from three treatment groups. d Bar plot depicting the cell percentages of each cell type as a proportion of total cells. Feature plot illustrating gene expression of ICOS ( e ) and CTLA-4 ( f ) in Foxp3 + Treg cells. g Bar plot depicting the cell percentages of ICOS high Treg cells within total CD3 + T cells. h – j MC38 tumor-bearing male mice were treated i.p. with CTR, 10 μg mCTLA4, or equimolar mProCTLA4 on day 11 after tumor inoculation. 2 days after the treatment, the total Treg cells and ICOS high Treg cells ( h ) from tumors were analyzed by flow cytometry. The frequency and absolute number of total Treg cells ( i , j ) and ICOS high Treg cells ( k , l ) was quantified. m Female Rag1 -/- mice ( n = 5/group) were i.v. transferred with Tregs. 3 days post-transfer, the mice were challenged s.c. with MC38-OVA. Activated OT-Ι T cells were transferred 4 days later. 5 days after OT-Ι transfer, mice were treated with 100 μg mProCTLA-4 every three days for three times, and the tumor curve was monitored. Data in panels ( b , i – m ) representative of two independent experiments. Statistical analysis was performed using ordinary one-way ANOVA with multiple comparisons test ( b, i – l ), or two-way ANOVA with Tukey’s multiple comparisons test ( m ).

Article Snippet: Cell types were predicted using SingleR (v.2.4.1) based on murine immune cell reference data (ImmGenData) from Celldex (v.1.12.0).

Techniques: Flow Cytometry, Isolation, Gene Expression