mouse ccr2 pe-conjugated antibody  (Bio-Techne corporation)

Journal: JCI Insight

Article Title: A prometabolite strategy inhibits cardiometabolic disease in an ApoE –/– murine model of atherosclerosis

doi: 10.1172/jci.insight.191090

Figure Lengend Snippet: ( A ) Schema of female ApoE –/– mice on an HFD with 150 mM SerBut or NaBut on a butyrate basis, or water ad libitum for 6 weeks. ( B ) Oil Red O (ORO) staining of aortic root plaque. ( C ) CD68 + IHC staining of aortic root plaque. Original magnification, ×20. ( D ) Representative images of immunofluorescent staining of aortic root. ( E ) Quantification of ORO + aortic root valve area. ( F ) Histological Stary scoring of plaque severity, ( G ) quantified CD68 + area, and ( H ) necrotic core of CD68 + IHC staining. ( I ) Quantified DAPI + , ( J ) DAPI + per plaque area, ( K ) iNOS + , and ( L ) iNOS + cells per plaque area in immunofluorescence stain. ( M – O ) Flow cytometry of immune cells infiltrating the aorta: ( M ) CD45 + total leukocytes, ( N ) CD11b + monocytes, and ( O ) CCR2 + CD11b + monocytes. ( P ) Water consumed by cages for the first 19 days of experiment. Data represent mean water consumed between 2 cages (replicates) as weight change of bottle converted to water volume. Error bars represent SD. ( Q ) Plasma IL-6 and ( R ) IFN-γ. ( S ) Blood flow cytometry of Ly6G + CD11b + circulating neutrophils. n = 5/cage and n = 10 mice per group. Data points represent individual mice displayed with median ± SEM. Statistical analyses were performed using a 1-way ANOVA with Tukey’s, Welch’s (if SDs were significantly different by Bartlett and Brown-Forsyth tests), or Kruskal-Wallis (if data were not normally distributed determined by Shapiro-Wilk test) post hoc test. P values less than 0.10 are shown. Outliers were removed by ROUT testing at Q = 1% in I – L . Scoring in C was done blinded. Data in C and F – H represent 2 independent pooled experiments: n = 10 for water and SerBut groups and n = 10 for NaBut groups in a single experiment. “B6,NC” denotes age-matched C57BL/6 mice on normal diet as non-statistical comparison to visualize healthy examples. Scale bars: 200 μm.

Article Snippet: In this study, antibodies used included those against CD16/CD32 (BioLegend, clone 93, catalog 101302), CCR2 (PE; R&D Systems, catalog FAB5538P), CD11b (BV605; BioLegend, clone M1/70, catalog 101257), CD11c (PE-Cy7; BD Biosciences, clone HL3, catalog 558079, BD Biosciences), CD19 (APC; BioLegend, clone 1D3, catalog 152412), CD3e (BUV395; clone 145-2C11, catalog 563565), CD45 (BV786; BD Biosciences, clone 30-F11, catalog 564225), F4/80 (BUV496; BD Biosciences, clone T45-2342, catalog 750644), I-A/I-E (PerCP-Cy5.5; BioLegend, clone M5/114.15.2, catalog 107626), Ly-6C (APC-Cy7; BioLegend, clone HK1.4, catalog 128026), and Ly-6G (FITC; eBioscience, clone 1AB, catalog 11-9668-80).

Techniques: Staining, Immunohistochemistry, Immunofluorescence, Flow Cytometry, Clinical Proteomics, Comparison

Journal: Frontiers in Immunology

Article Title: CD4 + skin resident memory T cells preferentially colocalize with dermal Folr2 hi macrophages in contact hypersensitivity

doi: 10.3389/fimmu.2025.1590687

Figure Lengend Snippet: Temporal changes in the number of skin myeloid cells after the resolution of CHS. (A, B) Representative flow cytometry plots of isolated naïve and healed ear skin cells on day 35 post-challenge. (A) CD11b + dermal myeloid cells were analyzed according to previous studies ( – ). CD11b + CD24 lo/- dermal myeloid cells in CD45 + Lineage - (Lin - : CD3ε - CD19 - Ly-6G - ) cells contained CD64 - Ly-6C - cells (cDC2) and remaining CCR2 + and CCR2 - populations. Based on Ly-6C and MHC-II expression, CCR2 + cells were divided into dermal monocytes (P1) and Ly-6C + (P2) and Ly-6C - (P3) MoDCs/CCR2 + macrophages. CCR2 - cells consisted of MHC-II lo/- (P4) and MHC-II hi (P5) macrophages. (B) DCs and LC populations were analyzed according to previous studies ( , ). CD45 + Lin - CD64 - Ly-6C - MHC-II hi cells consisted of CD24 + CD11b lo/- (cDC1), CD24 + CD11b + (LCs), CD24 - CD11b + (cDC2), and CD24 - CD11b - (DN cDCs). (C, D) The numbers of each cell lineage in naïve and healed ear skin were analyzed. (C) Absolute cell numbers were compared between days 37 and 318. (D) Temporal changes in the relative numbers of each myeloid cell population (expressed as a percentage relative to those in naïve skin) were analyzed at the indicated time points. Representative results are shown for experimental results conducted multiple times around days 37 and 100, while data from day 214 onward are from a single experiment. Data represent the mean ± S.E. (n = 5-6, each). * P < 0.05 [two-tailed paired (naïve vs healed) and unpaired t- test (days 37 vs 318)].

Article Snippet: The following antibodies were used for myeloid cell staining: anti-Ly-6C-FITC (HK1.4, BioLegend), CCR2-PE (475301, R&D systems), CD3ε-biotin (145-2C11, BD Biosciences, San Jose, CA), CD19-biotin (6D5, BioLegend), Ly-6G-biotin (1A8, BioLegend), CD24-PE-Cy7 (M1/69, BioLegend), Tim-4-PE-Cy7 (RMT4-54, BioLegend), Rat IgG2a/κ isotype control-PE-Cy7 (RTK2758, BioLegend), Folr2-APC (10/FR2, BioLegend), Rat IgG2a/κ isotype control-APC (RTK2758, BioLegend), CD45.2-APC-Vio770 (REA737, Miltenyi Biotec), MHC-II-BV421 (M5/114.15.2, BioLegend), CD64-BV605 (X54-5/7.1, BD Biosciences), and CD11b-BV786 (M1/70, BioLegend).

Techniques: Flow Cytometry, Isolation, Expressing, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: CD4 + skin resident memory T cells preferentially colocalize with dermal Folr2 hi macrophages in contact hypersensitivity

doi: 10.3389/fimmu.2025.1590687

Figure Lengend Snippet: Folr2 hi macrophages are the major macrophage population in skin. (A) The expression of common macrophage marker genes in dermal myeloid cell lineages in mouse naïve ear skin was obtained from a public database . The heat map shows microarray expression values (color scale based on percentiles). (B-F) The frequency of Folr2 hi cells in naïve and healed ear skin cells on day 38 was analyzed. Representative plots of CD45 and Folr2 expression in whole ear skin cells (B) , and Folr2 expression in P1-P5, Lin + (CD3ε + CD19 + Ly-6G + ), and other remaining CD45 + cell fractions (C) . (D) Pie charts show the distribution of total Folr2 hi cells to the indicated fractions. The size of pie charts represents the number of cells. “Other CD45 + ” and “stroma” fractions represent CD45 + cells other than the P3-P5 fractions and CD45 - cells, respectively. (E) Distribution of Folr2 hi and Folr2 lo/- cells in total P3-P5 fractions. (F) Numbers of Folr2 hi and Folr2 lo/- cells in P3-P5 fractions. Data are the mean ± S.E. (n = 5, for each). * P < 0.05 (naïve left ears vs healed right ears, two-tailed paired t- test). (G) Classification of skin macrophage populations proposed by Dick et al. and found in the present study. (H) Tim-4, Folr2, and MHC-II expression in the CD45 + Lin - CD11b + CD64 + CCR2 - macrophage population in naïve and healed ear skin on day 50. Graphs represent the numbers of MHC-II hi and MHC-I lo/- populations in the indicated fractions (mean ± S.E., n = 5, for each). P values (naïve left ears vs healed right ears, two-tailed paired t- test). (F, H) The macrophage subset included in each population is shown at the bottom of the graphs.

Article Snippet: The following antibodies were used for myeloid cell staining: anti-Ly-6C-FITC (HK1.4, BioLegend), CCR2-PE (475301, R&D systems), CD3ε-biotin (145-2C11, BD Biosciences, San Jose, CA), CD19-biotin (6D5, BioLegend), Ly-6G-biotin (1A8, BioLegend), CD24-PE-Cy7 (M1/69, BioLegend), Tim-4-PE-Cy7 (RMT4-54, BioLegend), Rat IgG2a/κ isotype control-PE-Cy7 (RTK2758, BioLegend), Folr2-APC (10/FR2, BioLegend), Rat IgG2a/κ isotype control-APC (RTK2758, BioLegend), CD45.2-APC-Vio770 (REA737, Miltenyi Biotec), MHC-II-BV421 (M5/114.15.2, BioLegend), CD64-BV605 (X54-5/7.1, BD Biosciences), and CD11b-BV786 (M1/70, BioLegend).

Techniques: Expressing, Marker, Microarray, Two Tailed Test