cluster differentiation 68 (cd68  (Agilent technologies)

Journal: Nature Communications

Article Title: Fibroblast growth factor signaling induces a chondrocyte-like state of peripheral nerve fibroblast during aging

doi: 10.1038/s41467-025-65297-8

Figure Lengend Snippet: a , b UMAP embedding of 1180 macrophage nuclei from sciatic nerves color-coded by subtype ( a ) or age ( b ). c Dot plot highlighting the marker genes used to identify the macrophage subtypes from A. Marker genes can be found in Table . d Relative percentage of Mϕs subtypes across ages. Steady state epi- and endoneurial Mϕ, circulating-derived phagocytic Mϕ, and chronic-inflammation Mϕ were more prevalent in sciatic nerves from 2-3, 15-16, and 20-30 months old mice, respectively. e Dot plot showing the expression levels of the marker genes used to identify the macrophage clusters, at the different time points. Markers for resident steady state Mϕ ( Cx3cr1, Mrc1, Csf1r ) are more expressed at 2-3 months old, those for circulating-derived phagocytic Mϕ (Cd74, Hspa5, Cd44, C1qa, Grn, Cd68), are more expressed at 15-16 months old, and those belonging to chronic inflammation ( Tgfβr1, Kynu) are more expressed in the 20-30 months old samples. f Representative immunofluorescence images of sciatic nerve samples show increased CD45 + /MRC1 + Mϕs in nerves from 2-3 months old mice, CD68 + /GRN + Mϕs in nerves from 15-16 and 20-30 months old mice, and CD45 + /TGF βR1+ in nerves from 20-30 months old mice. Yellow or pink arrowheads point to CD45 + /MRC1+ or CD68 + /GRN+ or CD45 + / TGF βR1 + cells. g Quantification of the number of CD45+ cells relative to the total number of cells. n = 7 - 9 - 10, biological replicates. h Quantification of the number of CD45 + /MRC1 + cells relative to the total number of CD45 + cells. n = 4 - 4 - 5 biological replicates. i Quantification of the number of CD68 + / GRN+ cells relative to the total number of CD68+ cells. n = 6 - 6 - 5. j Quantification of the number of CD45 + /TGF βR1 + cells relative to the total number of CD45 + cells. n = 3 - 5 - 5, biological replicates. Data are presented as mean values +/− SD. Source data for panels is provided as a Source Data file.

Article Snippet: Cluster of differentiation 68 (CD68) 1:250 , Mouse , MCA1957 Biorad Hercules, CA, USA.

Techniques: Marker, Derivative Assay, Expressing, Immunofluorescence

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: IL1RAP Expression in Human Atherosclerosis: A Target of Novel Antibodies to Reduce Vascular Inflammation and Adhesion

doi: 10.1161/JAHA.124.039557

Figure Lengend Snippet: A , Immunofluorescent staining depicts IL1RAP (green) alongside VWF (red), SMA (red), and the macrophage marker CD68 (red) within different areas of a human atherosclerotic plaque (in gray). All scalebars=100 μm except for whole plaque images, where the scalebar=1 mm. B , Expression of IL1R1 , IL1RAP , ST2 , and IL36R in human atherosclerotic plaques compared with donor‐matched macroscopically intact carotid tissue (n=32 biological replicates in each group), from the Gene Expression Omnibus database of microarray data. C , Expression of IL1R1, IL1RAP, ST2, and IL36R in human umbilical vein endothelial cells detected by flow cytometry shown as mean fluorescence intensity of fluorophore (APC, PE, A647)‐conjugated antibodies (Ab) towards the receptor components (IL1R1, IL1RAP, ST2, IL36R). CD68 indicates cluster of differentiation 68; DAPI, 4′,6‐diamidino‐2‐phenylindole; FC, fold change; IL‐1, interleukin 1; IL1R1, IL‐1 receptor 1; IL1RAP, interleukin 1 receptor associated protein; IL36R, IL‐36 receptor; SMA, smooth muscle actin; ST2, supression of tumorigenicity 2; and VWF, von Willebrand factor

Article Snippet: Cellular markers for endothelial cells (anti‐human von Willebrand factor; M0616; Dako, Denmark; 1:200), smooth muscle cells (anti‐human smooth muscle actin; M0851, Dako; 1:150), and macrophages (anti‐CD68 [cluster of differentiation 68]; Novocastra, Leica Biosystems, Newcastle, UK; 1:10) were used in parallel.

Techniques: Staining, Marker, Expressing, Gene Expression, Microarray, Flow Cytometry, Fluorescence

Journal: Molecular Neurodegeneration

Article Title: An adapted protocol to derive microglia from stem cells and its application in the study of CSF1R-related disorders

doi: 10.1186/s13024-024-00723-x

Figure Lengend Snippet: Functional phenotype of iMGL derived from the ALSP-CSF1R patient with a c.2350G > A (p.V784M) CSF1R variant. All iMGL were generated using the 2.9 protocol. Quantification ( A ) and images ( B ) of iMGL migratory activity toward ADP assessed by Boyden chamber assay. Cells were concomitantly treated or not with PSB0739. Kruskal–Wallis tests followed by Dunn’s multiple comparison tests were performed. n = 6 healthy control lines and 6 batches of a single ALSP patient line, differentiated side-by-side. White = Hoechst 33342, scale bar = 75 μm in ( B ). Quantification of green fluorescence intensity per cell ( C ) and representative images ( D ) of iMGL exposed to vehicle or pHrodo.™ Green-labelled myelin, opsonized red blood cells (IgG-RBC) or E. coli for three hours and then counterstained with Hoescht 33342. T-tests were performed. n = 3 healthy control lines and 3 batches of a single ALSP patient line, differentiated side-by-side, * p < 0.05. Scale bar = 250 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\mu m$$\end{document} μ m in ( D ). Quantification of mean fluorescence intensities (MFI; E ) and representative images ( F ) of LAMP1, CD68 and IBA1 immunostaining of iMGL. T-tests were performed. n = 3 healthy control lines and 3 batches of a single ALSP patient line, differentiated side-by-side, ** p < 0.01, *** p < 0.001. Scale bar = 100 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\mu m$$\end{document} μ m in ( F ). G Cytokine secretion assessed in cell supernatants following a 24-h treatment with vehicle, LPS (100 ng/mL) or Pam 3 CSK 4 (100 ng/mL). A two-way ANOVA was performed, followed by Tukey’s post hoc test. n = 4 healthy control lines and 4 batches of a single ALSP patient line, differentiated side-by-side, *** p < 0.001, ns = non-significant. H qRT-PCR performed after three hours of Pam 3 CSK 4 (100 ng/mL) vs. vehicle treatment. T-tests were performed. n = 3 healthy control lines and 3 batches of a single ALSP patient line, differentiated side-by-side, * p < 0.05

Article Snippet: Cells were incubated at 4 °C overnight with primary antibodies against the following targets: ionized calcium binding adaptor molecule 1 (IBA1; #NC9288364, Fujifilm Wako Chemicals at 1:1000), PU.1 (#2258, Cell Signaling at 1:250), Nanog (#ab21624, Abcam at 1:500), Tra-1–60 (#60064, STEMCell Technologies at 1:200), stage-specific embryonic antigen-4 (SSEA-4; #sc-21704, Santa Cruz Biotechnologies at 1:200), octamer-binding transcription factor 3/4 (OCT3/4; #sc-8628, Santa Cruz Biotechnologies at 1:500), Ki67 (#556003, BD Biosciences at 1:200), lysosomal-associated membrane protein 1 (LAMP1; #9091, Cell Signaling at 1:200) or cluster of differentiation 68 (CD68; #M0814, Dako Omnis at 1:200).

Techniques: Functional Assay, Derivative Assay, Variant Assay, Generated, Activity Assay, Boyden Chamber Assay, Comparison, Fluorescence, Immunostaining, Quantitative RT-PCR