mc38 mouse colon cancer cell line  (Absolute Biotech Inc)

Journal: Cell Reports Medicine

Article Title: Immunogenic tumor cell death and T-cell-derived IFN-γ elicit tumoricidal macrophages to potentiate OX40 immunotherapy

doi: 10.1016/j.xcrm.2026.102699

Figure Lengend Snippet: scRNA-seq reveals differential immune infiltration in αOX40-treated tumors based on response (A) Schematic of the bilateral MC38 tumor model assessing αOX40 response. Humanized OX40 mice received three doses of αOX40, followed by resection of the left tumor for scRNA-seq and flow cytometry analysis. Contralateral tumor dynamics and survival were monitored longitudinally. (B) UMAP visualization of scRNA-seq data from immune cells in MC38-bearing mice following αOX40 treatment. Cells are color-coded by annotated cell type. (C) Heatmap depicting nine transcriptionally distinct immune cell subpopulations. (D) Pie chart shows the relative abundance of nine immune cell clusters in αOX40 responders and nonresponders. (E) Flow cytometry analysis of tumor-infiltrating immune cell frequencies in αOX40-treated MC38-bearing mice. Frequencies of CD4 + T cells, CD8 + T cells, and macrophages were quantified after the third αOX40 dose (control, n = 5 mice; mice with a robust therapeutic response named as responder, n = 4 mice; mice with minimal to no response named as nonresponder, n = 4 mice). Data represent mean ± SD from one of two independent experiments (E). Statistical significance was determined using one-way ANOVA with multiple comparisons. ∗∗ p < 0.01.

Article Snippet: The mouse colon cancer MC38 cell line (Cat. NO. NM-S13) and the pancreatic cancer KPC cell line (Cat. NO. NM-YD04) were purchased from Shanghai Model Organisms Center, Inc.

Techniques: Flow Cytometry, Control, Clinical Proteomics

Journal: Cell Reports Medicine

Article Title: Immunogenic tumor cell death and T-cell-derived IFN-γ elicit tumoricidal macrophages to potentiate OX40 immunotherapy

doi: 10.1016/j.xcrm.2026.102699

Figure Lengend Snippet: NOS2-expressing macrophages is associated with response to αOX40 therapy (A) UMAP of monocytes/macrophages subclusters from scRNA-seq data in αOX40-treated MC38-bearing mice. (B) Representative marker genes in the monocyte/macrophage subclusters. (C) Pie chart showing the proportional distribution of monocyte/macrophage subsets of responders and nonresponders. (D) QuSAGE pathway analysis demonstrated enrichment of innate immune and phagocytic signaling pathways in distinct monocyte/macrophage subsets. (E) UMAP showing Mac_C1 signature genes and a heatmap of immune-related gene expression across TAM subclusters ( Z score normalized). (F) Violin plots comparing Nos2 expression levels in Mac_C1 subset between responsive and nonresponsive. (G) Flow cytometry analysis shows the percentage of M1-like (F4/80 + NOS2 + ) and M2-like (F4/80 + CD206 + ) macrophages in tumor tissues of control ( n = 5 mice), nonresponders (with minimal to no response, n = 4 mice), and responders (with a robust therapeutic response, n = 4 mice). (H and I) Comparison of Nos2 expression levels in responders versus nonresponders pre- or post-αOX40 treatment. Bilateral-MC38-bearing mice were treated with αOX40, and tumors from one side were analyzed by RNA-seq prior to (H) or following αOX40 treatment (I). The Nos2 expression was analyzed from RNA-seq data (left) and validated by RT-qPCR (right) ( n = 5 biological replicates). (J) NOS2 expression in tumor biopsies post-treatment determined by RNA-seq. Patients with advanced solid tumors and >1 prior therapy received HFB301001 monotherapy. Tumor biopsy samples were obtained on day 8 of cycle 2 for subsequent RNA-seq analysis. NOS2 expression were compared between patients achieving stable disease (SD, n = 3) and those with progressive disease (PD, n = 3). (K) GO enrichment analysis of upregulated genes in Mac_C1 of responders. (L) Calreticulin expression was quantified by flow cytometry in different response groups following αOX40 treatment ( n = 3 mice per group). (M) NOS2 expression in BMDMs was analyzed by flow cytometry after stimulation with CD8 + T cell supernatant and MC38 lysate, combined with TLR inhibition and IFN-γ blockade ( n = 5 biological replicates). (N) Quantification of Nos2 expression in BMDM by RT-qPCR after 24-h stimulation with MPLA (TLR4 agonist, 100 ng/mL), IFN-γ (20 ng/mL), or both. Data normalized to Gapdh and presented as fold-change relative to unstimulated controls ( n = 4 biological replicates). Data are shown as means ± SD from one of two independent experiments (G, H, I, L, M, and N). Statistical significance was determined using one-way ANOVA with multiple comparisons (G, L, M, and N) or using an unpaired two-tailed t test (H, I, and J). n.s., not significant; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001; VST, variance stabilized transformation; sup., supernatant; lys., tumor lysate; inh., inhibitor.

Article Snippet: The mouse colon cancer MC38 cell line (Cat. NO. NM-S13) and the pancreatic cancer KPC cell line (Cat. NO. NM-YD04) were purchased from Shanghai Model Organisms Center, Inc.

Techniques: Expressing, Marker, Protein-Protein interactions, Gene Expression, Flow Cytometry, Control, Clinical Proteomics, Comparison, RNA Sequencing, Quantitative RT-PCR, Inhibition, Two Tailed Test, Transformation Assay

Journal: Cell Reports Medicine

Article Title: Immunogenic tumor cell death and T-cell-derived IFN-γ elicit tumoricidal macrophages to potentiate OX40 immunotherapy

doi: 10.1016/j.xcrm.2026.102699

Figure Lengend Snippet: Rational modulation of tumor microenvironment enhances therapeutic responsiveness to αOX40-based immunotherapy (A–D) OX40-humanized mice bearing subcutaneous MC38 (A), B16 (B), E.G7 (C), or KPC (D) tumors ( n = 5–7 mice per group). Tumor growth curves (numbers indicate complete cures) and Kaplan-Meier survival for each model. Treatments: MPLA+IFN-γ ( i.t. , intratumoral); Combo: MPLA+IFN-γ ( i.t. , intratumoral) + αOX40 ( i.p. , intraperitoneal). (E) Study schema of secondary tumor challenge in MC38 model treated with Combo. (F) Tumor progression and survival outcomes following secondary tumor challenge. Growth kinetics of re-implanted tumors in tumor-cleared mice (previously cured by therapy) versus treatment-naive wild-type controls (left). Kaplan-Meier survival plot (right) ( n = 13 mice per group). (G) Systemic immunity evaluation schema with bilateral MC38 bearing mice were treated with Combo, αOX40, and control. (H) Tumor growth curves and survival plots of (G) ( n = 6–7 mice per group). Data are shown as means ± SD from one of two independent experiments (A–D, F, and H). Statistical significance was determined using log rank test (A–H). n.s., not significant; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: The mouse colon cancer MC38 cell line (Cat. NO. NM-S13) and the pancreatic cancer KPC cell line (Cat. NO. NM-YD04) were purchased from Shanghai Model Organisms Center, Inc.

Techniques: Control

Journal: Cell Reports Medicine

Article Title: Immunogenic tumor cell death and T-cell-derived IFN-γ elicit tumoricidal macrophages to potentiate OX40 immunotherapy

doi: 10.1016/j.xcrm.2026.102699

Figure Lengend Snippet: The antitumor efficacy of the Combo therapy is contingent upon CD8 + T cells and macrophages (A) UMAP of scRNA-seq data from tumor-infiltrating immune cells in OX40-humanized MC38-bearing mice treated with MPLA, IFN-γ, αOX40, or Combo. Cells are color-coded by annotated cell type. (B) Bubble chart showing the top variable marker genes for identified immune cell types. (C) Pie chart shows the relative abundance of 11 immune cell clusters in control, αOX40, MPLA+IFN-γ, or Combo. (D) Macrophage frequency and absolute count in tumors of MC38-bearing mice after two and three treatment cycles with MPLA, IFN-γ, αOX40, or Combo, analyzed by flow cytometry ( n = 5 mice per group). (E) Schematic of CD8 + T cell depletion assay. (F) Tumor volume and survival were monitored. Kaplan-Meier survival analysis corresponding to the depletion study of CD8 + T cell ( n = 6 mice per group). (G) Schematic of macrophage depletion assay in early and late stage. (H and I) Tumor volume and survival were monitored. Kaplan-Meier survival analysis corresponding to the depletion study in (G) ( n = 6–10 mice per group). Data are shown as means ± SD from one of two independent experiments (D, F, H, and I). Statistical significance was determined using one-way ANOVA with Tukey’s multiple comparisons test (D). Log rank test was used (F, H, and I) for statistical comparison. n.s., not significant; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: The mouse colon cancer MC38 cell line (Cat. NO. NM-S13) and the pancreatic cancer KPC cell line (Cat. NO. NM-YD04) were purchased from Shanghai Model Organisms Center, Inc.

Techniques: Marker, Control, Flow Cytometry, Depletion Assay, Comparison

Journal: Cell Reports Medicine

Article Title: Immunogenic tumor cell death and T-cell-derived IFN-γ elicit tumoricidal macrophages to potentiate OX40 immunotherapy

doi: 10.1016/j.xcrm.2026.102699

Figure Lengend Snippet: NOS2-high macrophages are significantly associated with Combo treatment efficacy (A) UMAP of macrophage subclusters from scRNA-seq data of MC38-bearing mice treated with control, MPLA+IFN-γ, αOX40, or Combo. Cells are color-coded by annotated subtype. (B) Bubble chart showing the top variable marker genes for identified macrophage subclusters. (C) Pie chart shows the relative abundance of four macrophage subclusters in control, αOX40, MPLA+IFN-γ, or Combo. (D) GO pathway analysis identifying significantly enriched signaling pathways in the Mac_S2 subcluster compared to other macrophage subpopulations. (E) Violin plots showing Nos2 expression levels across macrophage subclusters. (F) Violin plots comparing Nos2 and Cd206 expression levels among different treatment groups. (G) Frequency of M1-like, M2-like, or the ratio of M1/M2-like macrophage cells in tumor tissues from control, αOX40, MPLA+IFN-γ, and Combo groups with two time points, as determined by flow cytometry ( n = 5–10 mice per group). (H) Multiple immunofluorescence signal intensities of NOS2 + F4/80 + and CD206 + F4/80 + cells in the TME of control, αOX40, MPLA+IFN-γ, and Combo groups. Scale bars, 20 μm. Data are shown as means ± SD from one of two independent experiments (G and H). Statistical significance was determined using one-way ANOVA with Tukey’s multiple comparisons test (G). n.s., not significant; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: The mouse colon cancer MC38 cell line (Cat. NO. NM-S13) and the pancreatic cancer KPC cell line (Cat. NO. NM-YD04) were purchased from Shanghai Model Organisms Center, Inc.

Techniques: Control, Marker, Protein-Protein interactions, Expressing, Flow Cytometry, Immunofluorescence

Journal: Cell Reports Medicine

Article Title: Immunogenic tumor cell death and T-cell-derived IFN-γ elicit tumoricidal macrophages to potentiate OX40 immunotherapy

doi: 10.1016/j.xcrm.2026.102699

Figure Lengend Snippet: NOS2-dependent direct tumor cell killing by macrophages in Combo therapy (A) Schematic of co-culture using CFSE-labeled MC38 cells with tumor- or spleen-derived macrophages, with/without NIL treatment. (B) Flow cytometry quantification of 7-AAD + MC38 cells after 48 h co-culture with tumor-(right) or spleen (left)-derived macrophages ( n = 3 biological replicates). (C) Quantification of 7-AAD + MC38 cells after 48 h in vitro co-culture with or without NIL treatment ( n = 3 biological replicates). (D) Treatment schedule for MC38- or B16-tumor-bearing Nos2 KOor WT mice treated with Combo ( n = 6 mice per group). (E and F) Survival curves of MC38-bearing (E) and B16-bearing mice (F) were analyzed using the log rank test. (G) Flow cytometry analysis of 7-AAD + MC38 cells after 48-h co-culture with CFSE-labeled MC38 cells and BMDMs from Nos2 KO mice ( n = 5 biological replicates). (H) Phagocytosis rate of MC38 cells engulfed by BMDMs was assessed by flow cytometry ( n = 3 biological replicates). (I) Surface expression of CRT on MC38 cells was assessed by flow cytometry after co-culture with MPLA- and IFN-γ-polarized BMDMs in vitro ( n = 3 biological replicates). (J) Analysis of CALR + MC38 cells from MC38-tumor-bearing mice following final treatment with control or Combo, assessed by flow cytometry ( n = 5 biological replicates). Data are shown as means ± SD from one of two independent experiments (B, C, E, F, G, H, I, and J). Statistical significance was determined using one-way ANOVA with Tukey’s multiple comparisons (B, C, and I) or using unpaired Student’s t test (G, H, and J). Log rank tests (E and F) were also used for statistical analysis. n.s., not significant; ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Combo, MPLA, and IFN-γ combined with αOX86 (E and F).

Article Snippet: The mouse colon cancer MC38 cell line (Cat. NO. NM-S13) and the pancreatic cancer KPC cell line (Cat. NO. NM-YD04) were purchased from Shanghai Model Organisms Center, Inc.

Techniques: Co-Culture Assay, Labeling, Derivative Assay, Flow Cytometry, In Vitro, Expressing, Control

Journal: Cell Reports Medicine

Article Title: Immunogenic tumor cell death and T-cell-derived IFN-γ elicit tumoricidal macrophages to potentiate OX40 immunotherapy

doi: 10.1016/j.xcrm.2026.102699

Figure Lengend Snippet: Foxp3 + Treg depletion and macrophage reprogramming are involved in the anti-tumor effect of Combo (A) Flow cytometry analysis of TME. MC38-tumor-bearing mice were treated with control, αOX40, MPLA+IFN-γ, or Combo for two and three doses, and tumors were analyzed by flow cytometry. (B) Frequency and absolute count of CD25 + FOXP3 + cells in tumor tissues from control, αOX40, MPLA+IFN-γ, and Combo groups with two time points, as determined by flow cytometry ( n = 5 mice per group). (C) Treatment schedule for MC38-tumor-bearing Fcer1g KO or FcγRIIb KO mice. Mice were treated with Control, αOX40, MPLA+IFN-γ, and Combo every 3 days for a total of four doses. (D and E) Survival curves of Fcgr1g KO (D) and FcgrIIb KO (E) mice following treatment ( n = 5–6 mice per group) were monitored. (F) Treatment schedule. MC38-tumor-bearing mice were treated with MPLA and IFN-γ in combination with either OX40-mIgG2a or OX40-hIgG1 agonist antibodies (top), and the corresponding survival curves are shown (bottom) ( n = 5–7 mice per group). (G) Schematic of the co-culture experiment involving BMDMs and Tregs at a ratio of 1:4 (BMDM:Treg); Nos2 expression was measured by RT-qPCR. (H) Relative expression of Nos2 following the co-culture ( n = 4 biological replicates). (I) Multiple immunofluorescence (mIF) staining of MC38 tumors from mice treated with control, αOX40, MPLA+IFN-γ, or Combo, showing FOXP3 and NOS2 expression in border or intra-tumoral. Scale bars, 50 μm. (J) Analysis of cell numbers of FOXP3 and NOS2 expression at the border and intra-tumoral. Representative images from five randomly chosen fields were quantified with ImageJ. Data are shown as means ± SD from one of two independent experiments (B, D, E, F, H, and I). Statistical significance was determined using one-way ANOVA with Tukey’s multiple comparisons (B and H). Log rank test was also used (D–F). n.s., not significant; ∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Combo, MPLA, and IFN-γ combined with αOX86 (C–E).

Article Snippet: The mouse colon cancer MC38 cell line (Cat. NO. NM-S13) and the pancreatic cancer KPC cell line (Cat. NO. NM-YD04) were purchased from Shanghai Model Organisms Center, Inc.

Techniques: Flow Cytometry, Control, Co-Culture Assay, Expressing, Quantitative RT-PCR, Immunofluorescence, Staining

Journal: Nature

Article Title: Microbiota-driven antitumour immunity mediated by dendritic cell migration

doi: 10.1038/s41586-025-09249-8

Figure Lengend Snippet: a-g , GF mice transplanted with faeces from Resps or N-Resps were subcutaneously injected with MC38 tumour cells. Experimental scheme ( a ). Tumour growth curves ( b , left: summary, right: each mouse). n = 4–8 mice per group. c-g , The faecal samples were subjected to 16S rRNA gene sequencing to analyse the bacterial composition. The alpha diversity of the gut microbiota was scored by the Shannon index ( c ). Stacked bar plot of the family-level taxonomic composition of the faeces of Resp-FMT/N-Resp FMT mice. Only families with an abundance of >0.1% are shown ( d ). Differentially abundant taxa were analysed by LEfSe, the symbol sizes are scaled by relative abundance ( e ). The bar plot showed the LDA scores of differentially abundant taxa between the Resp and N-Resp groups ( f ). The relative abundance of Ruminococcaceae (left) and Bacteroidaceae (right)( g ). n = 14 mice per group. ( b , left), The average tumour sizes of the groups on a certain day of the experiment are shown as dots and are presented as the mean ± SD, two-way ANOVA with the Tukey—Kramer method. Each dot indicates one mouse, and the data are presented as the median ( c, g ). ( c , g ) Two-sided unpaired t test.

Article Snippet: The mouse colon cancer cell line MC38 (ENH204, Kerafast) and the mammary carcinoma cell line EMT6 (CRL-2755, ATCC) were cultured in high-glucose DMEM (Fujifilm Wako Pure Chemical) supplemented with 10% FBS (Biosera) and 2 mM l -glutamine.

Techniques: Injection, Sequencing

Journal: Nature

Article Title: Microbiota-driven antitumour immunity mediated by dendritic cell migration

doi: 10.1038/s41586-025-09249-8

Figure Lengend Snippet: a-l , ATB-SPF mice were transplanted with faeces from Resp or N-Resp mice following the subcutaneous injection of tumour cells. Experimental scheme ( a ). Tumour growth curves of MC38 ( b , left: summary, right: each mouse) and EMT6 ( c , left: summary, right: each mouse) tumour models. n = 5–6 mice per group. d-f , The frequency [representative staining (left) and % summary (right)] of effector molecule expression [PD-1 ( d , n = 10) and CD62L/CD44 ( e , n = 12)] and cytokine (IFN-γ/TNFα) production ( f , n = 12) by CD8 + T cells. g-i , TCR Vβ repertoire of PD-1 + CD8 + T cells ( g, h ). Numbers of skewed TCR Vβ clones (percentages of clones >10%) of PD-1 + CD8 + T cells ( i ). n = 12 mice per group. j-l , The expression [representative staining (left) and MFI summary (right)] of maturation markers [CD86 ( j ), CD40 ( k ) and CD80 ( l )] by DCs. n = 6 mice per group. ( b , left; c , left), The average tumour sizes of the groups on a certain day of the experiment are shown as dots and are presented as the mean ± SD. ( d-l ) Each dot indicates one mouse, and the data are presented as the mean ( i ) or mean ± SD ( d-f, h, j-l ). ( b, c ) Two-way ANOVA with the Tukey—Kramer method. ( d-f, j-l ), One-way ANOVA with Bonferroni correction. ( i) Two-sided Mann—Whitney U test.

Article Snippet: The mouse colon cancer cell line MC38 (ENH204, Kerafast) and the mammary carcinoma cell line EMT6 (CRL-2755, ATCC) were cultured in high-glucose DMEM (Fujifilm Wako Pure Chemical) supplemented with 10% FBS (Biosera) and 2 mM l -glutamine.

Techniques: Injection, Staining, Expressing, Clone Assay, MANN-WHITNEY

Journal: Nature

Article Title: Microbiota-driven antitumour immunity mediated by dendritic cell migration

doi: 10.1038/s41586-025-09249-8

Figure Lengend Snippet: a – e , The morphological traits of YB328. a , Left, scanning electron microscopy image. Scale bar, 1 µm. Right, transmission electron microscopy image of negatively stained cells. The arrows indicate extracellular membrane vesicles secreted by YB328. Scale bar, 0.5 µm. b , d , Relative abundances of YB328 ( b ) and P. vulgatus ( d ) in faecal samples of patients shown in Fig. ( n = 50). c , e , ROC curves (left) and Kaplan—Meier curves (right) of the PFS of patients with high or low abundances of YB328 ( c ) or P. vulgatus ( e ) in faeces. f , g , Tumour growth curves of SPF mice with B16F10 ( f ) or MC38 ( g ) tumours (left, summary; right, each mouse). n = 4–6 mice per group. Here and in other figures, control indicates mice treated with an isotype control antibody. h – j , The frequency (representative staining (left) and summary (right)) of PD-1 ( h , n = 8), CD62L and CD44 ( i , n = 7) and IFNγ and TNF production ( j , n = 7) by CD8 + T cells in MC38 tumours. Cells were gated on CD8 + TILs ( h – j ). k – m , The mean fluorescent intensity (MFI) of CD86 ( k , n = 6), CD40 ( l , control or anti-PD-1, n = 3, P. vulgatus or YB328, n = 4) and H-2Kb (a marker for MHCI) ( m , n = 6) by CD11c + MHCII + DCs in MC38 tumours. n – p , Tumour growth curves ( n , n = 5–7 mice per group), TCR Vβ repertoire ( o ) and numbers of skewed TCR Vβ clones (>10% of each clone) ( p ) among PD-1 + CD8 + T cells ( n = 5 mice per group). For b and d , each dot indicates one patient, and the data are presented as the median. For f (left), g (left) and n , the average tumour sizes of the groups on a certain day are shown as dots and are presented as the mean ± s.d. For h – m and p , each dot in the summary graphs indicates one mouse. Data are presented as the mean ± s.d. ( h – m ) or median ( p ). Significance was assessed using two-sided Mann—Whitney U -test ( b , d , p ), two-sided log-rank test ( c , right, e , right), two-way analysis of variance (ANOVA) with the Tukey—Kramer method ( f , g , n ) or one-way ANOVA with Bonferroni correction ( h – m ).

Article Snippet: The mouse colon cancer cell line MC38 (ENH204, Kerafast) and the mammary carcinoma cell line EMT6 (CRL-2755, ATCC) were cultured in high-glucose DMEM (Fujifilm Wako Pure Chemical) supplemented with 10% FBS (Biosera) and 2 mM l -glutamine.

Techniques: Electron Microscopy, Transmission Assay, Staining, Membrane, Control, Marker, Clone Assay, MANN-WHITNEY

Journal: Nature

Article Title: Microbiota-driven antitumour immunity mediated by dendritic cell migration

doi: 10.1038/s41586-025-09249-8

Figure Lengend Snippet: a , A representative gating strategy for evaluating the progeny of CD103 + CD11b − cDCs in BMDCs stimulated with YB328, P. vulgatus , or vehicle, corresponding to Fig. . b-c , Representative staining ( b ) of the phenotype of CD103 + CD11b − cDCs (% summary: c , left panel) or CD103 + DCs (% summary: c , right panel) in the indicated organs of WT or Batf3 −/− MC38 tumour-bearing mice (n = 3). d , Tumour growth curves (n = 6–9). e-f , ATB-SPF mice were subcutaneously injected with MC38 cells and administered an anti-PD-1 mAb, anti-CSF1R mAb or control Ab with/without YB328 treatment. Changes in the immunological landscape are shown ( e , n = 3). The experimental scheme ( f , upper panel) and tumour growth curves ( f , lower-left: summary, lower-right: each mouse) ( f , n = 4–7). g , OT-I CD8 + T cells were cocultured with bacteria-treated BMDCs (DC) or BMDMs (MF) pulsed with antigen peptides. The frequency of PD-1 expression [representative staining (left) and % summary (right)] by OT-I CD8 + T cells is shown (n = 5 biologically replicates). ( d ; f , lower-right) Each line indicates one mouse for the in vivo experiment. The average tumour size of the groups on a certain day is shown as a dot and are presented as the mean ± SD ( f , lower-left). Each dot in the summary graphs indicates one mouse in the in vivo experiment, and the data are presented as the mean ± SD ( c , e ). Each dot in the summary graphs indicates a well, and the data are presented as the mean ± SD ( g ). ( c, e ), Two-sided unpaired t test. ( f ) Two-way ANOVA with the Tukey—Kramer method. ( g ) One-way ANOVA with Bonferroni’s correction.

Article Snippet: The mouse colon cancer cell line MC38 (ENH204, Kerafast) and the mammary carcinoma cell line EMT6 (CRL-2755, ATCC) were cultured in high-glucose DMEM (Fujifilm Wako Pure Chemical) supplemented with 10% FBS (Biosera) and 2 mM l -glutamine.

Techniques: Staining, Injection, Control, Bacteria, Expressing, In Vivo

Journal: Nature

Article Title: Microbiota-driven antitumour immunity mediated by dendritic cell migration

doi: 10.1038/s41586-025-09249-8

Figure Lengend Snippet: a – f , ATB-SPF mice were subcutaneously injected with MC38 cells and treated with the indicated bacteria. The phenotype and frequency (representative staining (left) and frequency (right)) of DC populations in lamina propria (gated on CD11c + MHCII + cells) ( a ), Peyer’s patch (gated on CD11c + MHCII + cells) ( b ), dLNs (gated on live CD45 + cells) ( c ) and tumour (gated on CD11c + MHCII + cells) ( d ). Expression (representative staining (left) and MFI summary (right)) of CCR7 by CD103 + CD11b − cDCs in the MLNs ( e , n = 3). Expression (representative staining (left) and MFI summary (right)) of Ki67 by PD-1 + CD8 + T cells in dLNs and MLNs ( f , n = 5; gated on PD-1 + CD8 + CD3 + cells). g , Representative image (left) and staining (right) of KikG-expressing or KikR-expressing intestinal cells (gated on CD45 + cells). h , A representative image of the localization of migrated intestinal cells after the indicated bacterial treatment. i , The frequency of KikR + CD103 + CD11b − cDCs in CD45 + cells in dLNs and NdLNs. j , KikR + CD103 + DCs (representative staining (left) and number summary (right)) in tumours (per mg) (gated on CD45 + cells). k , The KikR − /KikR + ratio of PD-1 + CD8 + T cells (representative staining (left) and ratio summary (right)) in tumours. Cells were gated on PD-1 + CD8 + CD3 + cells. l , The TCR Vβ repertoire in CD8 + T cells in the indicated organs. For i–l , n = 3–6 mice per group. For a – l , each dot in the summary graphs indicates one mouse. The data are presented as the mean ± s.d. m – q , Assays in human samples. m , Representative multiplex immunohistochemical staining of tumours from patients with GC. Scale bars, 200 μm. n , o , Correlation between the YB328 abundance in the faeces of patients and the density of PD-1 + CD8 + T cells ( n ) or CLEC9A + IRF8 + cells ( o ). n = 20 patients. p , q , Correlation between PD-1 + CD8 + T cells and CLEC9A + IRF8 + cells ( p ) and correlation coefficients between PD-1 + CD8 + T cells and other immune cells ( q ). n = 24 patients. For n – p , each dot indicates one patient, and the relative correlation was assessed by two-sided Pearson’s correlation. Significance was assessed using two-sided unpaired Student’s t -test ( a – c , e , f , j – k ), one-way ANOVA with Bonferroni correction ( d , i ) or ANOSIM with one-tailed significance using 999 permutations ( l ).

Article Snippet: The mouse colon cancer cell line MC38 (ENH204, Kerafast) and the mammary carcinoma cell line EMT6 (CRL-2755, ATCC) were cultured in high-glucose DMEM (Fujifilm Wako Pure Chemical) supplemented with 10% FBS (Biosera) and 2 mM l -glutamine.

Techniques: Injection, Bacteria, Staining, Expressing, Multiplex Assay, Immunohistochemical staining, One-tailed Test

Journal: Nature

Article Title: Microbiota-driven antitumour immunity mediated by dendritic cell migration

doi: 10.1038/s41586-025-09249-8

Figure Lengend Snippet: a-c , ATB-SPF mice were subcutaneously injected with MC38 cells and treated with the indicated bacteria. The phenotype and frequency [representative staining (left) and % summary (right)] of DC populations in the indicated organs were examined: NdLNs ( a ) and SPs ( b ). n = 4–6 mice per group. TCF-1 expression by PD-1 + CD8 + T cells in the indicated organs after the indicated treatment. Representative staining (upper) and % summary (lower) ( c ). d-g , KikGR mice pretreated with ATB were injected subcutaneously with MC38 cells and treated with the indicated bacteria. Before bacterial treatment, KikGR mice were surgically photoconverted. Migratory intestinal cells that expressed KikR + were profiled in the indicated organs. The frequency [representative staining (left) and % summary (right)] of KikR + CD103 + CD11b − DCs ( d ), KikR + PD-1 + CD8 + T cells ( e ), KikR + Foxp3 + T cells ( f ) and KikR + CD11b + cells ( g ) among CD45 + cells. n = 3–6 mice per group. h , WT GF mice were subcutaneously injected with MC38 cells and were administered an anti-PD-1 mAb with YB328 treatment. Faeces were collected after the 24 h of the first administration of YB328. Tumour were collected at days 20. The presence of YB328 was examined. n.d., not detected. n = 7 mice per group. Each dot in the summary graphs indicates one mouse. All the data are presented as the mean ± SD. ( a , b , d-g ) Two-sided unpaired Student’s t test.

Article Snippet: The mouse colon cancer cell line MC38 (ENH204, Kerafast) and the mammary carcinoma cell line EMT6 (CRL-2755, ATCC) were cultured in high-glucose DMEM (Fujifilm Wako Pure Chemical) supplemented with 10% FBS (Biosera) and 2 mM l -glutamine.

Techniques: Injection, Bacteria, Staining, Expressing