Journal: Cell

Article Title: B cells and T follicular helper cells mediate response to checkpoint inhibitors in high mutation burden mouse models of breast cancer.

doi: 10.1016/j.cell.2019.10.028

Figure Lengend Snippet: (A) RNA-seq signatures for sensitive tumors at 7 days (5mm= day 0/ treatment initiation) without or with anti-PD1/anti-CTLA4 therapy. (B) Flow cytometry results for CD8+ cells and CD4+ using memory markers (Cd44, Cd62L). (C) Flow cytometry of tumor infiltrating B cells with or without ICI therapy. On the right shows staining for B cells gated for activation markers. (D) Quantification of flow cytometry for activated B cells (B220+, Cd19+ or Cd20+, MHC II +, Cd80+ or Cd86+). (E) IHC staining for IgG-kappa chain in KPB25Luv tumors. (F) IgG binding assay showing serum-IgG binding (Fitc+) to KPB25Luv cells. (G) Quantification of Fitc+ cells in IgG binding assay for KP25Luv cells and off-target binding. (H) Quantification of Fitc+ IgG binding assay for T11-Apobec cells following reabsorption on off-target cells. In boxplots, bars signify the mean and standard deviation. The p-values are two-tailed from unmatched T-tests. All tumors collected after 7days of treatment or non-treatment.

Article Snippet: Mouse IgG2b Isotype Control (clone MPC-11) , Bioxcel , Cat: BE0086; RRID: AB_1107791.

Techniques: RNA Sequencing, Flow Cytometry, Staining, Activation Assay, Immunohistochemistry, Binding Assay, Standard Deviation, Two Tailed Test

Journal: Cell

Article Title: B cells and T follicular helper cells mediate response to checkpoint inhibitors in high mutation burden mouse models of breast cancer.

doi: 10.1016/j.cell.2019.10.028

Figure Lengend Snippet: (A) Flow cytometry for B cells in KPB25Luv tumors after 7 days of ICI and CD4+ T cell depletion. (B) Quantification of results from A. (C) X-Y plot of IgG and CIBERSORT Tfh T cell signatures in mRNA-seq of sensitive tumors at day 7. (D) Boxplot of CIBERSORT Tfh T cell signature levels in sensitive tumors (mRNA-seq) at day 7. (E) X-Y plot of IgG signature and Il21 mRNA in mRNA-seq of sensitive tumors at day 7. (F) Boxplot of Il21 mRNA levels in sensitive tumors (mRNA-seq) at day 7. (G) Flow cytometry results for activated B cells in T11-Apobec & KPB25Luv tumors during Tfh/IL21 blockade. (H) IHC staining for IgG-kappa chain in KPB25Luv tumors during Tfh/IL21 blockade. (I) Survival for T11-Apobec bearing mice during ICI therapy and Tfh/IL21 blockade. (J) Survival for KPB25Luv bearing mice during ICI therapy and Tfh/IL21 blockade. (K) Western blot for serum IgG in Igmi and Balbc mice with T11-Apobec tumors. The blue bars mark Igmi mouse sera, purple note Balbc sera. (L) 21 day acute response in Igmi and Balbc control mice with T11-Apobec tumors. (M) Survival of Igmi mice withT11-Apobec tumors and treated with anti-PD1/anti-CTLA4 therapy in contrast to Balbc controls. (N) Survival in KPB25Luv tumor bearing mice treated with ICI therapy or ICI therapy with CD16/32 blockade . In Kaplan-Meier plots, p-values are from Log-rank (Mantel-Cox) tests. Boxplots show the mean and standard deviation. The p-values are two-tailed from standard T-tests. In X-Y plots, p-values were determined by linear regression analysis. The asterisks denote significance (***, p<0.0001, *, P<0.05).

Article Snippet: Mouse IgG2b Isotype Control (clone MPC-11) , Bioxcel , Cat: BE0086; RRID: AB_1107791.

Techniques: Flow Cytometry, Immunohistochemistry, Western Blot, Control, Standard Deviation, Two Tailed Test

Journal: Cell

Article Title: B cells and T follicular helper cells mediate response to checkpoint inhibitors in high mutation burden mouse models of breast cancer.

doi: 10.1016/j.cell.2019.10.028

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Mouse IgG2b Isotype Control (clone MPC-11) , Bioxcel , Cat: BE0086; RRID: AB_1107791.

Techniques: Control, Virus, Recombinant, Staining, Multiplex Assay, Enzyme-linked Immunosorbent Assay, Microarray, Plasmid Preparation, Software, Membrane, Transfection

Journal: Cell

Article Title: B cells and T follicular helper cells mediate response to checkpoint inhibitors in high mutation burden mouse models of breast cancer.

doi: 10.1016/j.cell.2019.10.028

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Mouse IgG2b Isotype Control (clone MPC-11) , Bioxcel , Cat: BE0086; RRID: AB_1107791.

Techniques: Control, Virus, Recombinant, Staining, Multiplex Assay, Enzyme-linked Immunosorbent Assay, Microarray, Plasmid Preparation, Software, Membrane, Transfection

Journal: Scientific Reports

Article Title: Next-generation Candida albicans vaccine VXV-01 containing recombinant Als3p and Hyr1p antigens for invasive Candida infections

doi: 10.1038/s41598-025-32488-8

Figure Lengend Snippet: Immunogenicity of Als3p/Hyr1p dual antigen vaccine formulated with alum, CAF01, or BDX100 adjuvants. (A). Combination of Als3p/Hyr1p doses by checkerboard method. (B). Schematic of the experimental design for determining vaccine-induced immunogenicity is shown. The ICR CD-1 mice ( N = 5/group) vaccinated SC (Alum, CAF01) or IN (BDX100) with different vaccine formulations (Als3p and Hyr1p antigens ratio on the x-axis) on days 0 and 21. Two weeks after the final vaccination, serum antigen-specific IgG titers and T cells were evaluated using ELISA and FluroSpot assay, respectively. Comparison of (C). Anti-Als3p and (D). anti-Hyr1p IgG endpoint titers in alum, CAF01 or BDX100 adjuvant-antigen formulations. Anti-Als3p and anti-Hyr1p IgG endpoint titers in (E). alum, (F). CAF01, (G). BDX100 adjuvant-antigen formulations. (H). Heat map showing the mean frequency ( n = 5 mice/cell/formulation) of Als3p or Hyr1p-specific Th1, Th2, and Th17 cells (IFN- γ, IL-4 or IL17 producing cells) in mice vaccinated with alum, CAF01 or BDX100 vaccine formulations. Each row represents data from each vaccine formulation. (I). Bar graph showing Als3p or Hyr1p-specific Th1, Th2, and Th17 cells.

Article Snippet: For CD4 + T cell depletion, 200 μg/mouse dose of rat anti-mouse CD4 IgG2b (clone GK1.5, BioXcell, Cat #BE0003-1)) or rat IgG2b isotype antibodies (Clone: LTF-2, BioXcell, Cat #BE0090) were administered intraperitoneally on day − 3 and 0 relative to infection.

Techniques: Immunopeptidomics, Enzyme-linked Immunosorbent Assay, Flurospot, Comparison, Adjuvant, Formulation

Journal: Scientific Reports

Article Title: Next-generation Candida albicans vaccine VXV-01 containing recombinant Als3p and Hyr1p antigens for invasive Candida infections

doi: 10.1038/s41598-025-32488-8

Figure Lengend Snippet: Mechanism of VXV-01-mediated protection. Naïve mice or vaccinated mice ( n = 10 mice/group) were infected with C. albicans or C. auris . For vaccination, the VXV-01 or placebo (adjuvant only) was administered on days 0, 21, and 35 (for C. albicans infection) or days 0 and 21 (for C. auris infection). Mice were immunosuppressed with cyclophosphamide and cortisone acetate for C. auris infection. Naïve mice ( n = 10 mice/group) infected with (A). C. albicans or (C). C. auris received two intraperitoneal anti-sera injections at 2 and 168 h relative to infection. Vaccinated mice ( n = 10 mice/group) received anti-CD4 or isotype control antibodies to deplete the CD T cells. The mice were infected with (B). C. albicans or (D). C. auris . Mice survivals were compared by Mantel-Cox test, and p < 0.5 was considered statistically significant.

Article Snippet: For CD4 + T cell depletion, 200 μg/mouse dose of rat anti-mouse CD4 IgG2b (clone GK1.5, BioXcell, Cat #BE0003-1)) or rat IgG2b isotype antibodies (Clone: LTF-2, BioXcell, Cat #BE0090) were administered intraperitoneally on day − 3 and 0 relative to infection.

Techniques: Infection, Adjuvant, Control

Journal: Scientific Reports

Article Title: Next-generation Candida albicans vaccine VXV-01 containing recombinant Als3p and Hyr1p antigens for invasive Candida infections

doi: 10.1038/s41598-025-32488-8

Figure Lengend Snippet: The durability of VXV-01 vaccine-induced immunity. The ICR (CD-1) mice were vaccinated sub-cutaneously with VXV-01 or placebo (adjuvant alone) on days: (A). 0, 21; or (B). 0, 21, 35. The serum anti-Als3p and anti-Hyr1p IgG endpoint titers and T cells were evaluated using ELISA and FluroSpot assay on days 14, 28, 90, 180, and 270 post-final vaccination. ( C ). Anti-Als3p, and ( D ). Hyr1p IgG endpoint titers and T-cell responses were compared between two and three vaccination schedules over the period of 270 days post-vaccination. Data presented as mean ± SE of N = 5 mice/group. ( E ). VXV-01 efficacy was tested after 15 weeks of final two doses vaccination regimen (day 0, 21) against C. auris infection in immunosuppressed mice ( N = 10 mice/group). Survival curves were compared by the Mantel-Cox test, and p < 0.5 was considered statistically significant.

Article Snippet: For CD4 + T cell depletion, 200 μg/mouse dose of rat anti-mouse CD4 IgG2b (clone GK1.5, BioXcell, Cat #BE0003-1)) or rat IgG2b isotype antibodies (Clone: LTF-2, BioXcell, Cat #BE0090) were administered intraperitoneally on day − 3 and 0 relative to infection.

Techniques: Adjuvant, Enzyme-linked Immunosorbent Assay, Flurospot, Infection

Journal: Nature Communications

Article Title: Munc13-4 mediates tumor immune evasion by regulating the sorting and secretion of PD-L1 via exosomes

doi: 10.1038/s41467-025-64149-9

Figure Lengend Snippet: A Western blot (WB) analysis of total PD-L1 level in control and Munc13-4 KO SUM159 or 4T1 cells (n = 3 biological replicates). B Representative TEM images of EVs secreted by control and Munc13-4 KO SUM159 or 4T1 cells (n = 3 biological replicates). Scale bar, 50 nm. C Quantification of exosomes secreted by equal numbers of control and Munc13-4 KO SUM159 (left) or 4T1 (right) cells through NTA (n = 3 biological replicates). D Schematic of experimental design for ( E , F ). WB analysis of PD-L1, Alix, CD63, and CD81 in EVs secreted by equal numbers of control and Munc13-4 KO SUM159 ( E ) or 4T1 ( F ) cell, collected from factions 1–6 (F1–6) (n = 3 biological replicates). WB analysis of PD-L1, Alix, CD63, and CD81 abundance on equal numbers of exosomes secreted by control and Munc13-4 KO SUM159 cells ( G ) and corresponding quantification of blot band intensities ( H ) (n = 4 biological replicates). WB analysis of PD-L1, Alix, CD63, and CD81 abundance on equal numbers of exosomes secreted by control and Munc13-4 KO 4T1 cells ( I ) and corresponding quantification of blot band intensities ( J ) (n = 3 biological replicates). K Schematic of experimental design for ( L ). L Quantification of killing efficiency against control and Munc13-4 KO 4T1 cells (n = 3 biological replicates). M Schematic of experimental design for ( N ). N Tumor growth curves following mammary gland inoculation of control or Munc13-4 KO 4T1 cells, with subsequent injection of PBS or the indicated exosomes (n = 6 mice). In ( A ), for each cell line, all samples were run on the same gel; in ( E , F , G , I ), samples were run on the same gel. Data are presented as means ± SEM ( C , H , J , N ), and p -values were calculated by unpaired two-tailed t-test ( C ), two-way ANOVA ( H , J ) with multiple comparisons, paired two-tailed t-test ( L ), and one-way ANOVA with multiple comparisons ( N ). Source data are provided as a file.

Article Snippet: Exosomes (1 × 10 9 particles) secreted by control 4T1 cells were pre-incubated with either an IgG isotype control antibody (Bioxcell, BE0090, 1:100) or InVivoMAb anti-mouse PD-L1 antibody (Bioxcell, BE0101, 1:100).

Techniques: Western Blot, Control, Injection, Two Tailed Test

Journal: Nature Communications

Article Title: Munc13-4 mediates tumor immune evasion by regulating the sorting and secretion of PD-L1 via exosomes

doi: 10.1038/s41467-025-64149-9

Figure Lengend Snippet: A Representative confocal images of SUM159 cells co-expressing GFP-PD-L1 with Orange-CD63 or Orange-LAMP1 (3 independent experiments). Scale bar, 20 μm. B Quantification of the mean Pearson’s correlation coefficient between PD-L1 and CD63 or LAMP1 (n = 3 independent experiments). C WB analysis of PD-L1, Alix, CD63, and CD81 on equal numbers of exosomes from SUM159 cells (n = 2 biological replicates). D WB analysis of total HRS in SUM159 cells (n = 3 biological replicates). E Co-IP/immunoblotting (IB) analysis in SUM159 cells transfected with indicated constructs (n = 3 biological replicates). F Representative confocal images of SUM159 cells in the PLA (3 independent experiments). Scale bar, 10 μm. G Quantification of the mean number of PLA puncta per cell (n = 3 independent experiments). H Co-IP/IB analysis in Munc13-4 KO SUM159 cells transfected with indicated constructs (n = 3 biological replicates). I Co-IP/IB analysis in HEK293T cells transfected with indicated constructs (n = 3 biological replicates). J Representative PLA image of SUM159 cells (3 independent experiments). Scale bar, 10 μm. K Co-IP/IB analysis in HEK293T cells transfected with indicated constructs (n = 3 biological replicates). L Representative PLA image of SUM159 cells (3 independent experiments). Scale bar, 10 μm. M Schematic of in vitro liposome co-flotation assay. N WB analysis of Munc13-4 and PD-L1 in top three fractions and bottom fraction (n = 3 biological replicates). O Co-IP/IB analysis in different SUM159 cells transfected with indicated constructs (n = 3 biological replicates). In ( C , D , N ), samples were run on the same gel; in ( E , H , I , K , O ), IP and Input samples were derived from the corresponding same experiment, but different gels for IP and Input samples were processed in parallel. Data are represented as means ± SEM ( B , G ), and p -values were calculated by two-way ANOVA with multiple comparisons ( B ) and unpaired two-tailed t-test ( G ). Source data are provided as a file.

Article Snippet: Exosomes (1 × 10 9 particles) secreted by control 4T1 cells were pre-incubated with either an IgG isotype control antibody (Bioxcell, BE0090, 1:100) or InVivoMAb anti-mouse PD-L1 antibody (Bioxcell, BE0101, 1:100).

Techniques: Expressing, Co-Immunoprecipitation Assay, Western Blot, Transfection, Construct, In Vitro, Derivative Assay, Two Tailed Test

Journal: Nature Communications

Article Title: Munc13-4 mediates tumor immune evasion by regulating the sorting and secretion of PD-L1 via exosomes

doi: 10.1038/s41467-025-64149-9

Figure Lengend Snippet: A Diagram of the sequences for P-pep and S-pep. P-pep comprises a cell-penetrating peptide (CPP) fused to the human PD-L1 256–273 motif, whereas S-pep consists of a CPP linked to a scrambled sequence containing the same amino acid composition as the human PD-L1 256–273 motif. B Co-IP/IB analysis in HEK293T cells transfected with indicated constructs and incubated with P-pep or S-pep to examine the effect of P-pep on Munc13-4–PD-L1 interaction (n = 3 biological replicates). C Co-IP/IB analysis in HEK293T cells transfected with indicated constructs and incubated with P-pep or S-pep to assess the effect of P-pep on the interactions of HRS with PD-L1 and Munc13-4 (n = 3 biological replicates). D WB analysis on the same number of exosomes secreted from equal number of SUM159 cells treated with S-pep or P-pep (n = 3 biological replicates). E Schematic of experimental design for the assessment of in vivo anti-tumor efficacy of P-pep. F Tumor growth curves of orthotopic mouse models of breast cancer treated with P-pep or S-pep (n = 9 mice). Flow cytometric quantification of the percentage of CD45 + CD3 + CD4 + ( G ) and CD45 + CD3 + CD8 + ( H ) T cells among total cells in tumors (n = 5 mice). Representative contour plots depicting CD45 + CD3 + CD4 + ( I ) and CD45 + CD3 + CD8 + ( K ) T cell populations within tumors, showing the expression of granzyme B (n = 5 mice). Quantification of the percentage of granzyme B + cells among CD45 + CD3 + CD4 + ( J ) and CD45 + CD3 + CD8 + ( L ) T cells within tumors (n = 5 mice). In ( B , C ), IP and Input samples were derived from the corresponding same experiment, but different gels for IP and Input samples were processed in parallel; in ( D ), all samples were run on the same gel. Data are represented as means ± SEM ( F ), box plots show the median (center line), interquartile range (box), minima and maxima (whiskers), and all individual data points (dots) ( G , H , J , L ), and p -values were all calculated by unpaired two-tailed t-test. Source data are provided as a file.

Article Snippet: Exosomes (1 × 10 9 particles) secreted by control 4T1 cells were pre-incubated with either an IgG isotype control antibody (Bioxcell, BE0090, 1:100) or InVivoMAb anti-mouse PD-L1 antibody (Bioxcell, BE0101, 1:100).

Techniques: Sequencing, Co-Immunoprecipitation Assay, Transfection, Construct, Incubation, In Vivo, Expressing, Derivative Assay, Two Tailed Test

Journal: Nature Communications

Article Title: Munc13-4 mediates tumor immune evasion by regulating the sorting and secretion of PD-L1 via exosomes

doi: 10.1038/s41467-025-64149-9

Figure Lengend Snippet: Schematic illustration showing that Munc13-4 collaborates with HRS, Rab27, and SNAREs to regulate PD-L1 sorting and secretion via exosomes. Loss of Munc13-4 in breast tumors enhances T cell-mediated anti-tumor immunity, suppresses tumor growth, and improves the efficacy of immune checkpoint inhibitors. Mechanistically, Munc13-4 regulates PD-L1 sorting by forming a ternary complex with PD-L1 and HRS. IFNγ stimulation modifies Munc13-4 and HRS, establishing a dynamic regulatory mechanism that enables tumor cells to adapt to immune pressure by modulating exosomal PD-L1 sorting. Downstream of sorting, Munc13-4 engages Rab27a to regulate MVB tethering and promotes SNARE complex assembly, thereby facilitating MVB–plasma membrane fusion and exosome release. Therapeutically, a designed peptide that disrupts the Munc13-4–PD-L1 interaction impairs PD-L1 sorting, resulting in enhanced anti-tumor immunity and reduced tumor growth in vivo.

Article Snippet: Exosomes (1 × 10 9 particles) secreted by control 4T1 cells were pre-incubated with either an IgG isotype control antibody (Bioxcell, BE0090, 1:100) or InVivoMAb anti-mouse PD-L1 antibody (Bioxcell, BE0101, 1:100).

Techniques: Clinical Proteomics, Membrane, In Vivo

Journal: Cell Death & Disease

Article Title: Targeting tumor intrinsic TAK1 engages TNF-α-driven cell death through distinct mechanisms and enhances cancer immunotherapy

doi: 10.1038/s41419-025-08013-0

Figure Lengend Snippet: A MC38 cells lacking Tak1 or B2m expression were pulsed with the MHC-I restricted chicken ovalbumin SIINFEKL peptide and subsequently cultured with increasing ratios of OT-I CD8 T cells, previously activated for 48 h by CD3 and CD28 stimulation. Tumor cell viability was assessed 24 h later by flow cytometry, using CD8 surface staining to discriminate T cells from tumor cells. B – D CD8 T cells isolated from naïve mouse spleens (Balb/c for CT-26 and EMT6, C57BL/6 for MC38) were activated by CD3 and CD28 stimulation in vitro and subsequently cultured with ( B ) MC38, ( C ) CT-26, or ( D ) EMT6 tumor cells lacking Tak1 expression. Viability was assessed using DRAQ7 uptake, imaging cells every 2 h for the indicated times. E – G Tak1- deficient MC38 ( E ) or CT-26 ( F ) cells were cultured in media derived from 48 h activated CD-8 T cells. Viability was assessed via DRAQ7 uptake. G Conditioned media from 48 h activated CD8 T cells was added to EMT6 cells deficient for Tak1 , Tnfr1 , or Tak1 and Tnfr1 and viability was assessed via DRAQ7 uptake. All panels represent the mean +/− SD from a single experiment, n = 2 independent experiments.

Article Snippet: For the CD8 depletion protocol, mice were injected with anti-mouse CD8 (BioXCell; BP0061, New Haven, CT) or IgG2a Isotype control (BioXCell; BP 00085) at 10 mg/kg i.p. at the dose schedule of Day -2, Day -1 prior to flank inoculation of cells, then again on Days 4, 8, and 11 post inoculation.

Techniques: Expressing, Cell Culture, Flow Cytometry, Staining, Isolation, In Vitro, Imaging, Derivative Assay

Journal: Cell Death & Disease

Article Title: Targeting tumor intrinsic TAK1 engages TNF-α-driven cell death through distinct mechanisms and enhances cancer immunotherapy

doi: 10.1038/s41419-025-08013-0

Figure Lengend Snippet: A Immunocompetent mice (C57BL/6, n = 15/group) were implanted with pooled clones from Tak1 -deficient or parental MC38 cells and tumor growth was monitored. B , C Immunocompetent mice (BALB/c, n = 8/group) were implanted with Tak1 -deficient CT-26 cells ( B ) and treated at the indicated days with an α-PD-1 antibody ( C , 10 mg/kg) and tumor growth was monitored. D Immunodeficient mice (NSG, n = 10/group) were implanted with Tak1 -deficient CT-26 cells and tumor growth was monitored. E Quantification of the percentage of complete tumor clearance from panels ( B – D ). F Balb/c mice implanted with Tak1 -deficient CT-26 tumors and exhibiting complete responses were rechallenged with CT-26 parental cells and tumor growth was monitored. Naïve mice challenged with parental CT-26 cells served as controls. G CT-26 parental and Tak1 -deficient tumor growth was monitored in Balb/c mice administered an α-CD8 depleting antibody for two sequential days prior to tumor engraftment and during tumor progression at the indicated days ( n = 10/group). H CT-26 parental and Tak1 -deficient tumor growth was assessed in Balb/c mice administered an α-TNF-α neutralizing antibody (15 mg/kg) at the indicated days ( n = 10/group). Tumor growth curves represent the mean tumor volume +/− SEM. Statistical differences between tumor volumes at the final measurement was determined using a two-way ANOVA with Tukey’s multiple correction where * p < 0.05; ** p < 0.01; and **** p < 0.0001 was considered significant.

Article Snippet: For the CD8 depletion protocol, mice were injected with anti-mouse CD8 (BioXCell; BP0061, New Haven, CT) or IgG2a Isotype control (BioXCell; BP 00085) at 10 mg/kg i.p. at the dose schedule of Day -2, Day -1 prior to flank inoculation of cells, then again on Days 4, 8, and 11 post inoculation.

Techniques: Clone Assay

Journal: Science Advances

Article Title: Immunosuppressive macrophages determine the effect of cellular senescence on tumor progression

doi: 10.1126/sciadv.adx2988

Figure Lengend Snippet: ( A ) Tumor samples from mice treated with vehicle or AP21967 at 9 weeks were homogenized, and the levels of the indicated cytokines were analyzed using a cytokine array. The log 2 FC (AP21967/vehicle) for each cytokine is presented. ENA-78, epithelial-derived neutrophil-activating peptide 78; GROα, growth-regulated oncogene-alpha. ( B ) Levels of CCL2 and CXCL1 were quantified by ELISA from the same tumor homogenates as in (A). ( C ) UMAP plots showing the expression of the most up-regulated cytokines (CCL2, CCL3, and CCL7) and their receptors (CCR1, CCR2, and CCR5) in different cell populations. Relevant cell clusters are indicated. N, neutrophils; MØ, macrophages; E, endothelial cells; F, fibroblasts. ( D ) Flow cytometry analysis of myeloid cells purified from tumors obtained from SuSe mice treated with vehicle or AP21967. We identified TAMs and cells positive for the myeloid marker CD11B and for the macrophage marker F4/80. ( E ) The same cells as in (D) were cultured for 24 hours—in the presence of vehicle or anti-CCL2—and the presence of CCL2 in the conditioned medium was determined by ELISA. ( F ) Schematic representation of the specificities of receptors for CCL2, CCL3, and CCL7. ( G ) Flow cytometry analysis of CCR2 in the same cells as in (D). FSH-H, forward scatter height.

Article Snippet: For selective CCL2 blocking, 4-week-old PyMT/SuSe mice were treated every 3 days with anti-CCL2 antibody (#BE0185, BioXCell,) at a concentration of 2 μg/g of body weight.

Techniques: Derivative Assay, Enzyme-linked Immunosorbent Assay, Expressing, Flow Cytometry, Purification, Marker, Cell Culture