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A) Experimental scheme for B16-OVA transplantation in nude mice followed by RMC-6236 oral gavage (10 mg/kg, daily) ; B) C omparison of tumor size between vehicle and RMC-6236–treated nude mice ; C) Tumor growth curves ; D,E) Flow-cytometry analysis of TAM CD206 (D) and CD86 (E) with vehicle or RMC-6236 ; F) Immunofluorescence showing macrophages <t>(F4/80,</t> green) and MHC-I (red) with DAPI (blue) ; G–I) Flow-cytometry quantification in TAMs of MHC-I (H-2K b ) (G) , OVA-specific presentation (SIINFEKL) (H) , and MHC-II (I-A/I-E) (I); J) Schematic of anti-CSF1R– mediated monocyte/macrophage depletion during RMC-6236 treatment; K) Tumor growth with RMC-6236 plus IgG or anti-CSF1R ; L, M) Intratumoral CD8 + T-cell frequency after monocyte depletion ; N,O) Granzyme B (N) and perforin (O) in tumor CD8 + T-cells. Data are shown as mean ± SEM. *p < 0.05, **p< 0.01, ***p < 0.001 and ****p < 0.0001 using unpaired two-tailed t-tests and Two-way ANOVA followed by Sidak’s multiple comparisons test (n≥6).
Anti Mouse F4 80 Microbeads, supplied by Miltenyi Biotec, 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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A) Experimental scheme for B16-OVA transplantation in nude mice followed by RMC-6236 oral gavage (10 mg/kg, daily) ; B) C omparison of tumor size between vehicle and RMC-6236–treated nude mice ; C) Tumor growth curves ; D,E) Flow-cytometry analysis of TAM CD206 (D) and CD86 (E) with vehicle or RMC-6236 ; F) Immunofluorescence showing macrophages <t>(F4/80,</t> green) and MHC-I (red) with DAPI (blue) ; G–I) Flow-cytometry quantification in TAMs of MHC-I (H-2K b ) (G) , OVA-specific presentation (SIINFEKL) (H) , and MHC-II (I-A/I-E) (I); J) Schematic of anti-CSF1R– mediated monocyte/macrophage depletion during RMC-6236 treatment; K) Tumor growth with RMC-6236 plus IgG or anti-CSF1R ; L, M) Intratumoral CD8 + T-cell frequency after monocyte depletion ; N,O) Granzyme B (N) and perforin (O) in tumor CD8 + T-cells. Data are shown as mean ± SEM. *p < 0.05, **p< 0.01, ***p < 0.001 and ****p < 0.0001 using unpaired two-tailed t-tests and Two-way ANOVA followed by Sidak’s multiple comparisons test (n≥6).
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A) Experimental scheme for B16-OVA transplantation in nude mice followed by RMC-6236 oral gavage (10 mg/kg, daily) ; B) C omparison of tumor size between vehicle and RMC-6236–treated nude mice ; C) Tumor growth curves ; D,E) Flow-cytometry analysis of TAM CD206 (D) and CD86 (E) with vehicle or RMC-6236 ; F) Immunofluorescence showing macrophages <t>(F4/80,</t> green) and MHC-I (red) with DAPI (blue) ; G–I) Flow-cytometry quantification in TAMs of MHC-I (H-2K b ) (G) , OVA-specific presentation (SIINFEKL) (H) , and MHC-II (I-A/I-E) (I); J) Schematic of anti-CSF1R– mediated monocyte/macrophage depletion during RMC-6236 treatment; K) Tumor growth with RMC-6236 plus IgG or anti-CSF1R ; L, M) Intratumoral CD8 + T-cell frequency after monocyte depletion ; N,O) Granzyme B (N) and perforin (O) in tumor CD8 + T-cells. Data are shown as mean ± SEM. *p < 0.05, **p< 0.01, ***p < 0.001 and ****p < 0.0001 using unpaired two-tailed t-tests and Two-way ANOVA followed by Sidak’s multiple comparisons test (n≥6).
F4/80 Antibody, Anti Mouse, Reafinity, supplied by Miltenyi Biotec, 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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A) Experimental scheme for B16-OVA transplantation in nude mice followed by RMC-6236 oral gavage (10 mg/kg, daily) ; B) C omparison of tumor size between vehicle and RMC-6236–treated nude mice ; C) Tumor growth curves ; D,E) Flow-cytometry analysis of TAM CD206 (D) and CD86 (E) with vehicle or RMC-6236 ; F) Immunofluorescence showing macrophages <t>(F4/80,</t> green) and MHC-I (red) with DAPI (blue) ; G–I) Flow-cytometry quantification in TAMs of MHC-I (H-2K b ) (G) , OVA-specific presentation (SIINFEKL) (H) , and MHC-II (I-A/I-E) (I); J) Schematic of anti-CSF1R– mediated monocyte/macrophage depletion during RMC-6236 treatment; K) Tumor growth with RMC-6236 plus IgG or anti-CSF1R ; L, M) Intratumoral CD8 + T-cell frequency after monocyte depletion ; N,O) Granzyme B (N) and perforin (O) in tumor CD8 + T-cells. Data are shown as mean ± SEM. *p < 0.05, **p< 0.01, ***p < 0.001 and ****p < 0.0001 using unpaired two-tailed t-tests and Two-way ANOVA followed by Sidak’s multiple comparisons test (n≥6).
Anti F4 80 Microbeads, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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A) Experimental scheme for B16-OVA transplantation in nude mice followed by RMC-6236 oral gavage (10 mg/kg, daily) ; B) C omparison of tumor size between vehicle and RMC-6236–treated nude mice ; C) Tumor growth curves ; D,E) Flow-cytometry analysis of TAM CD206 (D) and CD86 (E) with vehicle or RMC-6236 ; F) Immunofluorescence showing macrophages <t>(F4/80,</t> green) and MHC-I (red) with DAPI (blue) ; G–I) Flow-cytometry quantification in TAMs of MHC-I (H-2K b ) (G) , OVA-specific presentation (SIINFEKL) (H) , and MHC-II (I-A/I-E) (I); J) Schematic of anti-CSF1R– mediated monocyte/macrophage depletion during RMC-6236 treatment; K) Tumor growth with RMC-6236 plus IgG or anti-CSF1R ; L, M) Intratumoral CD8 + T-cell frequency after monocyte depletion ; N,O) Granzyme B (N) and perforin (O) in tumor CD8 + T-cells. Data are shown as mean ± SEM. *p < 0.05, **p< 0.01, ***p < 0.001 and ****p < 0.0001 using unpaired two-tailed t-tests and Two-way ANOVA followed by Sidak’s multiple comparisons test (n≥6).
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A) Experimental scheme for B16-OVA transplantation in nude mice followed by RMC-6236 oral gavage (10 mg/kg, daily) ; B) C omparison of tumor size between vehicle and RMC-6236–treated nude mice ; C) Tumor growth curves ; D,E) Flow-cytometry analysis of TAM CD206 (D) and CD86 (E) with vehicle or RMC-6236 ; F) Immunofluorescence showing macrophages <t>(F4/80,</t> green) and MHC-I (red) with DAPI (blue) ; G–I) Flow-cytometry quantification in TAMs of MHC-I (H-2K b ) (G) , OVA-specific presentation (SIINFEKL) (H) , and MHC-II (I-A/I-E) (I); J) Schematic of anti-CSF1R– mediated monocyte/macrophage depletion during RMC-6236 treatment; K) Tumor growth with RMC-6236 plus IgG or anti-CSF1R ; L, M) Intratumoral CD8 + T-cell frequency after monocyte depletion ; N,O) Granzyme B (N) and perforin (O) in tumor CD8 + T-cells. Data are shown as mean ± SEM. *p < 0.05, **p< 0.01, ***p < 0.001 and ****p < 0.0001 using unpaired two-tailed t-tests and Two-way ANOVA followed by Sidak’s multiple comparisons test (n≥6).
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A) Experimental scheme for B16-OVA transplantation in nude mice followed by RMC-6236 oral gavage (10 mg/kg, daily) ; B) C omparison of tumor size between vehicle and RMC-6236–treated nude mice ; C) Tumor growth curves ; D,E) Flow-cytometry analysis of TAM CD206 (D) and CD86 (E) with vehicle or RMC-6236 ; F) Immunofluorescence showing macrophages (F4/80, green) and MHC-I (red) with DAPI (blue) ; G–I) Flow-cytometry quantification in TAMs of MHC-I (H-2K b ) (G) , OVA-specific presentation (SIINFEKL) (H) , and MHC-II (I-A/I-E) (I); J) Schematic of anti-CSF1R– mediated monocyte/macrophage depletion during RMC-6236 treatment; K) Tumor growth with RMC-6236 plus IgG or anti-CSF1R ; L, M) Intratumoral CD8 + T-cell frequency after monocyte depletion ; N,O) Granzyme B (N) and perforin (O) in tumor CD8 + T-cells. Data are shown as mean ± SEM. *p < 0.05, **p< 0.01, ***p < 0.001 and ****p < 0.0001 using unpaired two-tailed t-tests and Two-way ANOVA followed by Sidak’s multiple comparisons test (n≥6).

Journal: bioRxiv

Article Title: Macrophage Antigen Presentation Is Unleashed by Pan-RAS Inhibition to Promote Antitumor Immunity

doi: 10.64898/2026.01.30.702886

Figure Lengend Snippet: A) Experimental scheme for B16-OVA transplantation in nude mice followed by RMC-6236 oral gavage (10 mg/kg, daily) ; B) C omparison of tumor size between vehicle and RMC-6236–treated nude mice ; C) Tumor growth curves ; D,E) Flow-cytometry analysis of TAM CD206 (D) and CD86 (E) with vehicle or RMC-6236 ; F) Immunofluorescence showing macrophages (F4/80, green) and MHC-I (red) with DAPI (blue) ; G–I) Flow-cytometry quantification in TAMs of MHC-I (H-2K b ) (G) , OVA-specific presentation (SIINFEKL) (H) , and MHC-II (I-A/I-E) (I); J) Schematic of anti-CSF1R– mediated monocyte/macrophage depletion during RMC-6236 treatment; K) Tumor growth with RMC-6236 plus IgG or anti-CSF1R ; L, M) Intratumoral CD8 + T-cell frequency after monocyte depletion ; N,O) Granzyme B (N) and perforin (O) in tumor CD8 + T-cells. Data are shown as mean ± SEM. *p < 0.05, **p< 0.01, ***p < 0.001 and ****p < 0.0001 using unpaired two-tailed t-tests and Two-way ANOVA followed by Sidak’s multiple comparisons test (n≥6).

Article Snippet: Macrophages were purified using anti-mouse F4/80 MicroBeads (Miltenyi Biotec) according to the manufacturer’s protocol.

Techniques: Transplantation Assay, Flow Cytometry, Immunofluorescence, Two Tailed Test