Journal: Frontiers in Immunology

Article Title: Activated hepatic stellate cell-derived small extracellular vesicles facilitate M2 macrophage polarization and hepatoma progression via miR-27a-3p

doi: 10.3389/fimmu.2024.1489679

Figure Lengend Snippet: miR-27a-3p promotes the M2 polarization of macrophage. (A) Representative images for the immunohistochemical staining of macrophage maker CD68 and CD206 in nude mice xenografts, bar = 100 μm; and the number of CD68, CD206 positive cells per 200 × field for each group (mean ± SEM). **P < 0.01, ***P < 0.001 vs control. Below each group of images is an image showing the magnification of the corresponding box area above. (B) The expression of miR-27a-3p in mimic-miR-27a-3p or anti-miR-27a-3p treated THP1 cells was evaluated by qRT-PCR 24 h after transfection, normalized to U6 snRNA (mean ± SEM). **P < 0.01, ****P < 0.0001 vs. control. (C, D) The expression of CD206 was evaluated in mimic-miR-27a-3p or anti-miR-27a-3p treated THP1 cells by western blotting normalized to β-actin (mean ± SEM). *P < 0.05, ****P < 0.0001 vs. control. The data were from three independent experiments.

Article Snippet: Monoclonal antibodies against CD206 (Boster, China), CD68 (Boster, China), Ki67 (Abcam, USA), and SPRY2 (Proteintech, China) were used.

Techniques: Immunohistochemical staining, Staining, Control, Expressing, Quantitative RT-PCR, Transfection, Western Blot

Journal: eLife

Article Title: Human-induced pluripotent stem cell-derived microglia integrate into mouse retina and recapitulate features of endogenous microglia

doi: 10.7554/eLife.90695

Figure Lengend Snippet:

Article Snippet: Antibody , anti-human CD68 (mouse monoclonal) , R&D , Cat. #: MAB20401 , IHC (1:100).

Techniques: Virus, Recombinant, In Situ, Plasmid Preparation, cDNA Synthesis, SYBR Green Assay, Flow Cytometry, Staining, Lysis, Bicinchoninic Acid Protein Assay, Software

Journal: bioRxiv

Article Title: Transglutaminase 2 function in glioblastoma tumor efferocytosis

doi: 10.1101/2024.08.29.610293

Figure Lengend Snippet: A and B. TGM2 RNA expression in glioblastoma molecular subtypes in the TCGA and CGGA datasets; C and D. Correlation of TGM2 mRNA levels with mRNA levels for the macrophage/microglia marker CD68 in the TCGA and CGGA datasets; E. The fifty genes showing the highest positive Spearman correlation with TGM2 mRNA expression in the TCGA database were identified using cBioPortal. This gene set was then analyzed for matches to cell type datasets. Yellow highlights show the match to macrophage cell type identified using Enrichr under Cell Types/ARCHS4 Tissues (34/50 match, qval = 2.8×10-15, odds ratio 5.35); F. TGM2 mRNA expression in single cell RNAseq data. Plots show UMAP clustering using data from Abdelfattah et al . analyzed with the Broad Institute Single Cell Portal. Plots show cell type assignments (left) and TGM2 mRNA expression (right); G. Violin plots (with all data points) of TGM2 mRNA expression in glioblastoma tumour cell types; H. Violin plots (with all data points) of TGM2 mRNA in the glioblastoma tumour immune cell subtypes described by Abdelfattah et al.

Article Snippet: Anti-Iba1/AIF1 mouse monoclonal antibody (used for immunofluorescence in xenografts) and anti-CD68 mouse monoclonal antibody (used for immunofluorescence in human tissue samples) were from Millipore/Sigma (cat. #s MABN92 and AMAB90873, respectively).

Techniques: RNA Expression, Marker, Expressing

Journal: bioRxiv

Article Title: Transglutaminase 2 function in glioblastoma tumor efferocytosis

doi: 10.1101/2024.08.29.610293

Figure Lengend Snippet: A. Image from full section showing strong TGM2 expression near regions of necrosis (indicated with N). Regions without necrosis (lower left) show lower levels of TGM2 staining; B. Necrotic region showing TGM2 positive cells surrounding the border of the necrotic region, similar to what was observed in the xenograft model. C-F. Double immunofluorescence for CD68 and TGM2 in whole sections from patient tumours. For each image row, the left panel shows CD68, the middle panel shows TGM2, and the right panel shows the merged image. Except for E, each row shows a single focal plane image from the Z stack. Row C shows a region with macrophages and endothelial cells. Row D shows a higher magnification view of a tumour blood vessel with adjacent macrophages to illustrate differences in subcellular location. Row E shows a Z stack of the same region as B, emphasizing the co-localization of TGM2 and CD68 in macrophages. Row F shows an area adjacent to necrotic regions.

Article Snippet: Anti-Iba1/AIF1 mouse monoclonal antibody (used for immunofluorescence in xenografts) and anti-CD68 mouse monoclonal antibody (used for immunofluorescence in human tissue samples) were from Millipore/Sigma (cat. #s MABN92 and AMAB90873, respectively).

Techniques: Expressing, Staining, Immunofluorescence

Journal: Cell Reports Methods

Article Title: A co-culture system of macrophages with breast cancer tumoroids to study cell interactions and therapeutic responses

doi: 10.1016/j.crmeth.2024.100792

Figure Lengend Snippet: Purification, freezing, and thawing of PBMCs from blood (A) PBMCs were purified by a classical Ficoll gradient followed by differentiation into macrophages using M-CSF for 7 days. Representative immunofluorescence images depict CD68 staining after differentiation. A control image was included without the primary antibody. (B) Experimental procedure outlining the thawing of PBMCs and their differentiation into macrophages. (C) Thawed PBMCs were differentiated into macrophages with M-CSF for 7 days. Representative immunofluorescence images illustrate CD68 staining after differentiation. A control image was included without the primary antibody. (D) Flow cytometry results comparing the cell viability of macrophages after using two different post-thaw wash media. (E) Flow cytometry results comparing the viability of macrophages after detachment from the plate using two different detachment media.

Article Snippet: Monoclonal Mouse anti-human CD68 (1/50) , Santa Cruz Biotechnology , Cat# sc-17832;RRID: AB_627157.

Techniques: Purification, Immunofluorescence, Staining, Control, Flow Cytometry

Journal: Cell Reports Methods

Article Title: A co-culture system of macrophages with breast cancer tumoroids to study cell interactions and therapeutic responses

doi: 10.1016/j.crmeth.2024.100792

Figure Lengend Snippet: Light-sheet microscopy of transparized tumoroids to characterize macrophage infiltration in the three setups (A) Illustration of the experimental procedure of the immunofluorescence and clearing protocol of the three models. (B) Representative light-sheet microscopy images of the tumoroids with CytoTell green-stained macrophages in the three models. (C) Representative light-sheet microscopy images of the tumoroids with CD68-labeled macrophages (green) in the three models.

Article Snippet: Monoclonal Mouse anti-human CD68 (1/50) , Santa Cruz Biotechnology , Cat# sc-17832;RRID: AB_627157.

Techniques: Microscopy, Immunofluorescence, Staining, Labeling

Journal: Cell Reports Methods

Article Title: A co-culture system of macrophages with breast cancer tumoroids to study cell interactions and therapeutic responses

doi: 10.1016/j.crmeth.2024.100792

Figure Lengend Snippet:

Article Snippet: Monoclonal Mouse anti-human CD68 (1/50) , Santa Cruz Biotechnology , Cat# sc-17832;RRID: AB_627157.

Techniques: Control, Recombinant, Cell Culture, Mass Spectrometry, Viability Assay, Staining, Software