Journal: Translational Oncology

Article Title: Vaccine-induced ErbB (EGFR/HER2)-specific immunity in spontaneous canine cancer

doi: 10.1016/j.tranon.2021.101205

Figure Lengend Snippet: cEGFR p527 immunization mediates CD8 infiltration and antibody deposition into canine tumors. Postmortem bladder cancer tissue from a cEGFR p527 immune dog and tissue from healthy, non-cancerous bladder was examined for infiltration of CD8 T cells and endogenous antibody. A. CD8 isotype control. B. CD8 T cell staining (green). Arrows illustrate presence of CD8+ cells within the tumor tissue. C. & D. Antibody deposition in canine bladder tumor tissue from a cEGFR p527 immunized dog (two different fields). E. & F. Canine IgG deposition in normal canine bladder tissue (two different fields). Magnification is 40x (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.).

Article Snippet: CD8 T cells were stained in tissues using mouse anti-dog CD8 (MAB6709; R&D Systems, Minneapolis, MN), followed by goat anti-mouse IgG ( H + L ) Alexa Fluor 488 (A11017; Life Technologies, Eugene, OR) and counterstaining with DAPI (Molecular Probes; Eugene, OR).

Techniques: Control, Staining

Journal: Science China. Life sciences

Article Title: Total body irradiation of donors can alter the course of tolerance and induce acute rejection in a spontaneous tolerance rat liver transplantation model.

doi: 10.1007/s11427-012-4370-3

Figure Lengend Snippet: Figure 3 Detection of T cell subpopulations in the peripheral blood and CTL-mediated lytic activity. A, There was no difference in the percentage of CD3+ T and CD3+CD4+ T cells between the transplant groups. The number of CD3+CD8+ T cells in the acute rejection group increased significantly (P<0.05), while the number of CD4+/CD8+ cells decreased significantly compared to the other groups (P<0.05). B, The ratio of Foxp3+CD4+CD25+ Tregs and the expression of CD4+CD25+ T cells in the acute rejection group was significantly lower than those in the other groups (P<0.05). C, In the control group, GITR expression was not detectable on T cells. GITR expression on increased T cells from all groups after culture with phorbol ester and ionomycin. The GITR expression level of T cells in the acute rejection group was significantly higher than that in the other groups (P<0.05). D, The spleen CTL-mediated lytic activity of the recipients in the acute rejection group increased by 32.6%, and was significantly higher than those in the other three groups (P<0.05).

Article Snippet: A 50 μL sample of the whole blood obtained after stimulation was incubated with 10 μL of anti-CD3-PerCp and 5 μL of anti-CD8-APC-Cy7 mAbs in the dark at 4°C for 30 min. After haematolysis and washing with PBS, the resuspended cells were incubated in the dark at 4°C for 45 min with 10 μL anti-rat GITR-PE mAb (R&D Systems, Minneapolis, MN, USA) or 10 μL IgG1-PE (Beckman Coulter Inc) as an isotype-control.

Techniques: Activity Assay, Expressing, Control

Journal: Nature communications

Article Title: Monoacylglycerol lipase regulates cannabinoid receptor 2-dependent macrophage activation and cancer progression.

doi: 10.1038/s41467-018-04999-8

Figure Lengend Snippet: Fig. 2 Macrophage MGLL inhibits tumor progression via CD8+T cells. a Six-week-old WT and TgMGLL mice were subcutaneously inoculated with MC-38 cells, and tumor volumes were measured dynamically. This test was performed four times. (n = 10) b The survival time of the MC-38 tumor-bearing mice described in a. The data represent the means ± s.e.ms. (n = 10, ***P < 0.005, Gehan-Breslow-Wilcoxon test). c The growth curves of the subcutaneous MC-38 tumors in Myeloid-mgll-KO mice and the control littermates (fl/fl). (n = 6). d The survival time of the MC-38 tumor-bearing mice described in c. The data represent the means ± s.e.ms. (n = 6, **P < 0.01, Gehan-Breslow-Wilcoxon test). e Incidence of metastasis in lungs and livers of the intravenous tumor model. The 6-week-old C57BL/6 WT or transgenic mice were intravenously injected with MC-38 colorectal cancer cells (5 × 106 cells per mouse) via the tail vein. The mice were sacrificed 2 weeks after tumor inoculation. The lungs and livers were dissected for pathological observation of tumor lesions. Data are means ± s.e.ms. and polled from four individual experiments. Each symbol represents an experimental replicate (n = 8~11). f–h The MC-38 tumor lesions in lungs of WT and TgMGLL mice, as described in e. Representative images of gross anatomy (f) and H&E staining (g) are displayed. The blue dotted lines indicate the tumor lesions. The relative tumor lesions were calculated (h). Scale bars, 200 μm. (n = 8~9). i Macrophage MGLL inhibited tumor growth in a Rag-1 dependent mannaer. MC-38 tumors were inoculated subcutaneously in WT, TgMGLL, Rag1KO or TgMGLL + Rag1KO mice and the tumor volumes were measured dynamically. (n = 5) j mRNA levels of IFNγ in CD8+ or CD4+ T cells that were isolated from the MC-38 tumors inoculated subcutaneously in WT or TgMGLL mice for 3 weeks. (n = 5). k Six-week-old WT or TgMGLL mice were subcutaneously inoculated with MC-38 cells and treated with anti-CD8 antibody or IgG as a control. The tumor volume was measured dynamically. (n = 10). l The survival time of the MC-38 tumor- bearing mice treated as described in k. The data represent the means ± s.e.ms. (n = 10, *P < 0.05, Gehan-Breslow-Wilcoxon test; ns, not significant). Data in a, c and e–k represent the means ± s.e.ms. (*P < 0.05, **P < 0.01, ***P < 0.005; student’s t-test; ns not significant)

Article Snippet: The MC-38 tumor-bearing mice were treated with monoclonal antibodies anti-CD8 (MAB116, Novus Biologicals) or IgG (MAB002, Novus Biologicals) as control.

Techniques: Control, Transgenic Assay, Injection, Staining, Isolation

Journal: Nature communications

Article Title: Monoacylglycerol lipase regulates cannabinoid receptor 2-dependent macrophage activation and cancer progression.

doi: 10.1038/s41467-018-04999-8

Figure Lengend Snippet: Fig. 6 MGLL-CB2 axis regulates tumor progression in mice. a Relative mRNA levels of TNFα, IL-6 and IFNγ in the CD8+ T cells in the MC-38 tumors inoculated subcutaneously in WT, TgMGLL, TgCB2, and TgMGLL+CB2 mice. The data represent the means ± s.e.ms. (n = 5, ***P < 0.005, Student’s t-test; ns not significant). b Six-week-old WT, TgMGLL, TgCB2, and TgMGLL+CB2 mice were subcutaneously inoculated with MC-38 cells, and the tumor volume was measured dynamically. This experiment was repeated three times. The data represent the means ± s.e.ms. (n = 10, **P < 0.01, ***P < 0.005, Student’s t-test). c The survival time of the MC-38 tumor-bearing mice. The mice were described in b. The data represent the means ± s.e.ms. (n = 10, **P < 0.01, ***P < 0.005, Gehan-Breslow-Wilcoxon test). d The incidence of tumor metastasis in MC-38 tumor model. Six-week-old WT, TgMGLL, TgCB2, and TgMGLL

Article Snippet: The MC-38 tumor-bearing mice were treated with monoclonal antibodies anti-CD8 (MAB116, Novus Biologicals) or IgG (MAB002, Novus Biologicals) as control.

Techniques:

Journal: Life (Basel, Switzerland)

Article Title: Diets Differently Regulate Pulmonary Pathogenesis and Immune Signaling in Mice during Acute and Chronic Mycobacterium tuberculosis Infection.

doi: 10.3390/life13010228

Figure Lengend Snippet: Figure 3. Representative immunohistochemistry (IHC) images of the lung sections of mice during acute Mtb infection (30 DPI) stained by CD4+, CD8+, F4/80+, and IFNγ antibodies. (RD_con— uninfected mice fed an RD; MFD_con—uninfected mice fed an MFD; RD_Inf—infected mice fed an RD; and MFD_Inf—infected mice fed an MFD). Magnification-4×; Scale bar–100 µ. IHC sections were graded and presented as a bar graph (see Supplementary Figure S3a).

Article Snippet: The blots were probed with the following primary antibodies: PPARα-specific mouse monoclonal antibody (#MA1-822, ThermoFisher Scientific), Perilipin 1-specific rabbit monoclonal antibody (#9349, Cell Signaling Technology), Phospho-Perilipin 1 (Ser522)-specific mouse monoclonal antibody (#4856, Vala Sciences), F4/80-specific rat monoclonal (#NB600-404, Novus Biologicals), adiponectinspecific mouse monoclonal antibody (#ab22554, Abcam), IFNγ-specific rabbit monoclonal antibody (#ab133566, Abcam), TNFα-specific rabbit polyclonal antibody (#ab6671, Abcam), CD4 specific rabbit polyclonal antibody (#NB100-56457SS, Novus Biologicals), CD8 specific rabbit polyclonal antibody (#NBP2-29475, Novus Biologicals), BNIP3 specific rabbit polyclonal antibody (#44060, Cell signaling Technology), Caspase-3-specific rabbit polyclonal antibody (#9662, Cell signaling Technology), IL-6-specific mouse monoclonal antibody (#66146-1-Ig, Proteintech), and IL-10-specific mouse monoclonal antibody (#60269-1-Ig, Proteintech).

Techniques: Immunohistochemistry, Infection, Staining

Journal: Life (Basel, Switzerland)

Article Title: Diets Differently Regulate Pulmonary Pathogenesis and Immune Signaling in Mice during Acute and Chronic Mycobacterium tuberculosis Infection.

doi: 10.3390/life13010228

Figure Lengend Snippet: Figure 4. MFD alters the level of infiltrated immune cells and inflammatory markers in the lungs during chronic Mtb infection. Immunoblot analysis of (a) immune cells (CD4+, CD8+, and F4/80+) and (b) inflammatory markers (IFNγ, TNFα, and IL-6) in the lung lysates of RD-fed and MFD-fed uninfected and infected C57BL/6 mice during chronic (90 DPI) infection. The error bars represent standard error of the mean. * p < 0.05, ** p < 0.01, and *** p < 0.001 between indicated groups.

Article Snippet: The blots were probed with the following primary antibodies: PPARα-specific mouse monoclonal antibody (#MA1-822, ThermoFisher Scientific), Perilipin 1-specific rabbit monoclonal antibody (#9349, Cell Signaling Technology), Phospho-Perilipin 1 (Ser522)-specific mouse monoclonal antibody (#4856, Vala Sciences), F4/80-specific rat monoclonal (#NB600-404, Novus Biologicals), adiponectinspecific mouse monoclonal antibody (#ab22554, Abcam), IFNγ-specific rabbit monoclonal antibody (#ab133566, Abcam), TNFα-specific rabbit polyclonal antibody (#ab6671, Abcam), CD4 specific rabbit polyclonal antibody (#NB100-56457SS, Novus Biologicals), CD8 specific rabbit polyclonal antibody (#NBP2-29475, Novus Biologicals), BNIP3 specific rabbit polyclonal antibody (#44060, Cell signaling Technology), Caspase-3-specific rabbit polyclonal antibody (#9662, Cell signaling Technology), IL-6-specific mouse monoclonal antibody (#66146-1-Ig, Proteintech), and IL-10-specific mouse monoclonal antibody (#60269-1-Ig, Proteintech).

Techniques: Infection, Western Blot

Journal: Life (Basel, Switzerland)

Article Title: Diets Differently Regulate Pulmonary Pathogenesis and Immune Signaling in Mice during Acute and Chronic Mycobacterium tuberculosis Infection.

doi: 10.3390/life13010228

Figure Lengend Snippet: Figure 6. MFD alters immune signaling in adipose tissue during (a) acute and (b) chronic Mtb infection. Immunoblot analysis of immune cells (CD4, CD8, and F4/80) and inflammatory markers (IFNγ, TNFα, IL-6, and IL-10) in the WAT lysates of RD-fed and MFD-fed uninfected and infected C57BL/6 mice during acute infection (30 DPI) and chronic (3 months) post-infection is shown. The error bars represent standard error of the mean. * p < 0.05, ** p < 0.01, and *** p < 0.001 between indicated groups.

Article Snippet: The blots were probed with the following primary antibodies: PPARα-specific mouse monoclonal antibody (#MA1-822, ThermoFisher Scientific), Perilipin 1-specific rabbit monoclonal antibody (#9349, Cell Signaling Technology), Phospho-Perilipin 1 (Ser522)-specific mouse monoclonal antibody (#4856, Vala Sciences), F4/80-specific rat monoclonal (#NB600-404, Novus Biologicals), adiponectinspecific mouse monoclonal antibody (#ab22554, Abcam), IFNγ-specific rabbit monoclonal antibody (#ab133566, Abcam), TNFα-specific rabbit polyclonal antibody (#ab6671, Abcam), CD4 specific rabbit polyclonal antibody (#NB100-56457SS, Novus Biologicals), CD8 specific rabbit polyclonal antibody (#NBP2-29475, Novus Biologicals), BNIP3 specific rabbit polyclonal antibody (#44060, Cell signaling Technology), Caspase-3-specific rabbit polyclonal antibody (#9662, Cell signaling Technology), IL-6-specific mouse monoclonal antibody (#66146-1-Ig, Proteintech), and IL-10-specific mouse monoclonal antibody (#60269-1-Ig, Proteintech).

Techniques: Infection, Western Blot

Journal: Cell Death & Disease

Article Title: Oncogenic activation of SMYD3-SHCBP1 promotes breast cancer development and is coupled with resistance to immune therapy

doi: 10.1038/s41419-025-07570-8

Figure Lengend Snippet: a The protein levels of Smyd3 and Shcbp1 in normal mouse mammary glands, and tumors initiated with HP5712 cells in 3 months old FVB virgin mice as shown by Western blots. b The protein levels of Smyd3 and Shcbp1 in HP5712 cells expressing sgSmyd3, OE-Smyd3, or OE-Smyd3/sgShcbp1 by Western blots. Representative tumor images ( c ) and tumor weight plots ( d ) at day 32 from FVB virgin mice implanted with HP5712 parental (HP tumors), OE-Smyd3-HP5712, sgSmyd3-HP5712, and OE-Smyd3/sgShcbp1-HP5712 cells at 1X10 6 cells per mammary fat pad (n = 8 mice/group). e The plot of relative spleen weight from the same cohort of mice in c and d . f The protein levels of Smyd3, H3K4me3, and Shcbp1 in tumors initiated with HP5712 parental, OE-Smyd3-HP5712, sgSmyd3-HP5712, and OE-Smyd3/sgShcbp1-HP5712 cells by Western blots. g The protein levels of pMek, pErk, and Kras in tumors initiated with HP5712 parental, OE-Smyd3-HP5712, sgSmyd3-HP5712, and OE-Smyd3/sgShcbp1-HP5712 cells as shown by Western blots. h tSNE visualized immune cells from normal FVB mouse mammary glands (FVB MG, n = 6 mice), tumors implanted with parental HP5712 cells (HP Vector Ctr), HP-OE-Smyd3, HP-OE-Smyd3/sgShcbp1, and HP-sgSmyd3 from FVB mice (n = 6 mice/group) by CyTOF analysis. Quantifications of PMN-MDSCs and M-MDSCs ( i ), CD4+ and CD8 + T cells ( j ) in CD45+ immune cell populations from the same cohorts of mice in h (n = 3 biological independent samples/per group). k , l The WT-T cells were activated with CD3 and CD28 antibody. Co-cultured the activated T cells without or with MDSCs from the mice implanted with HP parental cells, HP cells expressing Smyd3, or sgSmyd3, or OE-Smyd3/sgShcbp1 for 72 hours and examined T cells proliferation status ( k ) and quantifications ( l ) of data in k (n = 3 mice/group). Representative images of CD8+/PD1+ double positive cells in mammary tissues from the mice with the genotypes of HP-Ctr, HP-OE-Smyd3, HP-sgSmyd3, and HP-OE-Smyd3/sgShcbp1 ( m ) and quantifications of CD8+/PD1 double positive cells from mammary tissues ( n ) in these mice (n = 4 mice/group, and at least 20 images for each sample were counted). Scale bar: 20 μm. arrow heads point to PD1/CD8 positive cell. o Expressions of PD1 from the spleen with implantation of parental HP cells, HP cells with OE-Smyd3, HP-sgSmyd3, and HP-OE-Smyd3/sgShcbp1 in FVB mice as determined by qPCR. p Summary of Smyd3-Shcbp1oncogenic signals shape the TIME.

Article Snippet: In Vivo anti-mouse CD8a , BioX Cell , BP0117 , , .

Techniques: Western Blot, Expressing, Plasmid Preparation, Cell Culture

Journal: Cell Death & Disease

Article Title: Oncogenic activation of SMYD3-SHCBP1 promotes breast cancer development and is coupled with resistance to immune therapy

doi: 10.1038/s41419-025-07570-8

Figure Lengend Snippet: Representative tumor images ( a ), and tumor weight plots ( b ) at day 22 from FVB mice implanted with HP5712 cells at 1×10 6 cells per mammary fat pad, and treatment with PBS, PD1, trametinib (Tra), and PD1+Tra (n = 8 mice/group). c The plot of tumor volume in the processes of treatment in ( a , b ). d The plot of relative spleen weight from the same cohort of mice in a , b . Quantifications of PMN-MDSCs and M-MDSCs ( e ), CD4+ and CD8 + T cells ( f ) in CD45+ immune cell populations from the same cohorts of mice in a , b by CyTOF analysis at D12 (n = 3 mice/group). Quantifications of PMN-MDSCs and M-MDSCs ( g ), CD4+ and CD8 + T cells ( h ) in CD45+ immune cell populations from the same cohorts of mice in a , b by CyTOF analysis at D22 (n = 3 mice/group). i , j Tumor images from the HP Ctr mice, HP mice treated with αPD1 and Tra, and HP mice treated with αPD1 and Tra with depletion of T cell using CD8 antibody ( i ). The plot of tumor weight ( j ) in the same cohort of mice in ( i ) (n = 8 mice in each group). The protein levels of Smyd3 and Shcbp1 in tumors initiated with HP5712 cells and treatment with PBS, αPD1, Tra, and αPD1+Tra in FVB mice at D12 ( k ) and D22 ( l ) as shown by Western blots. The protein levels of pMek, pErk, Kras, and Grb2 in tumors initiated with HP5712 cells and treatment with PBS, αPD1, Tra, and αPD1+Tra in FVB mice at D12 ( m ) and D22 ( n ) as shown by Western blots. Expressions of Smyd3 ( o ) and Shcbp1 ( p ) in G600 cells with the treatment of E2, Tra, and E2 together with Tra at 1 hour, 2 hours, 4 hours, and 24 hours as determined by qPCR.

Article Snippet: In Vivo anti-mouse CD8a , BioX Cell , BP0117 , , .

Techniques: Western Blot

Journal: Cell Death & Disease

Article Title: Oncogenic activation of SMYD3-SHCBP1 promotes breast cancer development and is coupled with resistance to immune therapy

doi: 10.1038/s41419-025-07570-8

Figure Lengend Snippet: Antibodies used for ChIP, IHC, IF, WB and in vivo mouse study.

Article Snippet: In Vivo anti-mouse CD8a , BioX Cell , BP0117 , , .

Techniques: Immunohistochemistry-IF, In Vivo

Journal: Journal of experimental & clinical cancer research : CR

Article Title: Regnase-1 downregulation promotes pancreatic cancer through myeloid-derived suppressor cell-mediated evasion of anticancer immunity.

doi: 10.1186/s13046-023-02831-w

Figure Lengend Snippet: Fig. 6 Suppression of CTLs by CD11b+ MDSCs is responsible for the acceleration of tumor progression by Regnase-1 downregulation. A-D Evaluation of phenotypes of orthotopic syngeneic tumors of WT or Regnase-1 KO murine pancreatic cancer cells. Representative macro images of pancreatic tumors (A). Relative mRNA levels of Cd8a, Ifng, Fasl, and Gzmb (B) (N = 6 per group). Representative images of HE (C, left panel) and CD8a immunostaining (C, right panel) and the number of CD8-positive cells (C, right) (N = 6 per group). Dot plots of CD3+CD8+ cells evaluated by flow cytometry (D, left) and the proportion of CD8 + cells among CD45+ cells (D, right) (N = 3 per group). E–H Evaluation of phenotypes of orthotopic syngeneic tumors of WT or Regnase-1-KO murine pancreatic cancer cells with or without depletion of CD8+ cells upon anti-CD8a antibody or IgG treatment. Experimental schematic (E). Dot plots of CD3+ and CD8.+ cells in WT or Regnase-1-KO syngeneic tumors upon anti-CD8a antibody or IgG treatment evaluated by flow cytometry (F). Tumor weights (G) (N = 6 per group). The relative mRNA levels of Cd8a, Ifng, Fasl, and Gzmb (H) (N = 6 per group). Student’s t test was used to evaluate differences between 2 groups. One-way ANOVA with Tukey’s post hoc test was used to compare differences among 4 groups. *P < 0.05, scale bars: 100 μm (insets)

Article Snippet: BE0061, a fully neutralizing mAb recognizing CD8a, and control IgG were obtained from Bioxcell.

Techniques: Immunostaining, Flow Cytometry

Journal: NPJ Precision Oncology

Article Title: Clock pathway inhibitor overcomes tumor immune-exclusion via regulation of fibrocyte differentiation

doi: 10.1038/s41698-025-01066-6

Figure Lengend Snippet: A Double staining of CD8 and αSMA in mouse various subcutaneous tumors. Scale bar, 200 μm. The quantitative evaluation of sections from each tumor stained for B αSMA ( n = 12 fields from four mice per group) and C CD8 ( n = 12 fields from four mice per group). D The correlation between the number of CD8 + T cells and αSMA + area in each mouse tumor tissue. E Double staining of collagen 1 and CD45 to detect fibrocytes in mouse various subcutaneous tumors. Scale bar, 100 μm. F The quantitative evaluation of sections from each tumor stained for fibrocytes ( n = 12 fields from four mice per group). G The correlation between the number of fibrocytes and αSMA + area in each mouse tumor tissue. The correlation was estimated by Spearman’s correlation and a linear regression analysis (the best-fit line is indicated). Representative images of tumor sections resected from non-small cell lung carcinoma (NSCLC) patients stained to detect αSMA + cancer-associated fibroblasts ( H , αSMA + FAP + ) and fibrocytes ( I , CD45 + FSP-1 + ). Scale bar, 100 μm. J Quantitative evaluation of tumor-infiltrating fibrocytes (CD45 + FSP-1 + ) in NSCLC tumor tissue stained in Fig. 1H and I ( n = 50 patients). K Representative images of resected NSCLC tissue stained to detect CD8 + T cells and EpCAM + tumor cells. Scale bar, 100 μm. L Quantitative evaluation of tumor-infiltrating fibrocytes (CD45 + FSP-1 + ) in each NSCLC group divided by the immune phenotypes of tumor-infiltrating CD8 + T cells (inflamed, desert, and exclusion). * P < 0.05 by a one-way ANOVA. All data are shown as the mean ± s.e.m.

Article Snippet: CD8 + and CD4 + T cells, macrophages, and Tregs were identified using a rat anti-CD8a monoclonal antibody (1:150, 53-6.7; BD Pharmingen, Franklin Lakes, NJ, USA), rat anti-mouse CD4 (1:150, H129.19; BD Pharmingen), rat anti-mouse F4/80 monoclonal antibody (1:100 dilution, CI:A3-1; Abcam, Cambridge, MA, USA), and rabbit anti-Foxp3 polyclonal antibody (1:400 dilution; Novus Biologicals, Centennial, CO, USA), respectively.

Techniques: Double Staining, Staining

Journal: NPJ Precision Oncology

Article Title: Clock pathway inhibitor overcomes tumor immune-exclusion via regulation of fibrocyte differentiation

doi: 10.1038/s41698-025-01066-6

Figure Lengend Snippet: A The evaluation of the tumor volume of AB1-HA-bearing balb/c nude mice treated with KL001 from 5 days after tumor cell injection ( n = 6 per group). B Double staining of CD8 and αSMA in AB1-HA tumors from balb/c mice treated with or without KL001 (studied in Fig. ). Scale bar, 200 μm. C Representative images and D quantitative evaluation of sections from KL001-treated AB1-HA tumors stained for CD8, CD4, Foxp3, and F4/80 ( n = 15 fields from five mice per group). Scale bar, 200 μm. E Quantitative evaluation of sections from CLK8-treated AB1-HA tumors stained for CD8, CD4, Foxp3 ( n = 15 fields from five mice per group). * P < 0.05 by the Mann-Whitney U test. All data are shown as the mean ± s.e.m.

Article Snippet: CD8 + and CD4 + T cells, macrophages, and Tregs were identified using a rat anti-CD8a monoclonal antibody (1:150, 53-6.7; BD Pharmingen, Franklin Lakes, NJ, USA), rat anti-mouse CD4 (1:150, H129.19; BD Pharmingen), rat anti-mouse F4/80 monoclonal antibody (1:100 dilution, CI:A3-1; Abcam, Cambridge, MA, USA), and rabbit anti-Foxp3 polyclonal antibody (1:400 dilution; Novus Biologicals, Centennial, CO, USA), respectively.

Techniques: Injection, Double Staining, Staining, MANN-WHITNEY

Journal: NPJ Precision Oncology

Article Title: Clock pathway inhibitor overcomes tumor immune-exclusion via regulation of fibrocyte differentiation

doi: 10.1038/s41698-025-01066-6

Figure Lengend Snippet: A The evaluation of the tumor volume and B the representative image of tumor tissue of LLC-bearing mice treated with KL001 and/or αPD-L1 Ab (n = 7 per group). * P < 0.05 by a one-way ANOVA. C Representative images and D the quantitative evaluation of sections from LLC tumors stained for αSMA ( n = 15 fields from five mice per group). The tumors were harvested at day 21 from each group studied in Fig. 6A. Scale bar, 200 μm. Representative images and the quantitative evaluation of sections from LLC tumors stained for collagen 1 + CD45 + fibrocytes ( E, F ), CD8 + T cells ( G, H ) and CD4 + T cells ( I, J ). Scale bar, 100 μm. * P < 0.05 by a one-way ANOVA. K The evaluation of the tumor volume of AB1-HA-bearing mice treated with KL001 and/or αCTLA-4 Ab ( n = 6 per group). * P < 0.05 by a one-way ANOVA. The quantitative evaluation of sections from AB1-HA tumors ( n = 15 fields from five mice per group) stained for CD8 ( L ), CD4 ( M ), and Foxp3 ( N ). The tumors were harvested at day 18 from each group studied in Fig. 6K. * P < 0.05 by a one-way ANOVA. All data are shown as the mean ± s.e.m.

Article Snippet: CD8 + and CD4 + T cells, macrophages, and Tregs were identified using a rat anti-CD8a monoclonal antibody (1:150, 53-6.7; BD Pharmingen, Franklin Lakes, NJ, USA), rat anti-mouse CD4 (1:150, H129.19; BD Pharmingen), rat anti-mouse F4/80 monoclonal antibody (1:100 dilution, CI:A3-1; Abcam, Cambridge, MA, USA), and rabbit anti-Foxp3 polyclonal antibody (1:400 dilution; Novus Biologicals, Centennial, CO, USA), respectively.

Techniques: Staining