Journal: Frontiers in Immunology

Article Title: Unveiling spatial complexity in solid tumor immune microenvironments through multiplexed imaging

doi: 10.3389/fimmu.2024.1383932

Figure Lengend Snippet: Immunophenotyping panel for multiplexed tissue imaging of cancer.

Article Snippet: CD206 , DCN228 , 50 , 130-123-671 , FITC (PE) , Miltenyi Biotec.

Techniques: Imaging

Journal: Frontiers in Immunology

Article Title: Unveiling spatial complexity in solid tumor immune microenvironments through multiplexed imaging

doi: 10.3389/fimmu.2024.1383932

Figure Lengend Snippet: Deep spatial profiling of human palatine tonsil tissues. (A) Hematoxylin and eosin (H&E) staining after 92 MICS cycles, including the marked epithelium, germinal center (GC), and T cell zone of the lymphoid follicle. MICS DAPI and stroma staining depicting the composition and structure of the tonsil. Markers: Collagen III, collagen IV, fibronectin (all extracellular matrix (ECM), cytokeratin (epithelium), podoplanin (lymphatic vessels), CD105/SM Actin (blood vessels). (B) Immune cell content of a human palatine tonsil comprising T cells (CD3), B cells (CD19/CD20), plasma cells (PCs) (CD38/CD138), NK cells (CD56), granulocytes (CD15/CD66b), mast cells (CD117), macrophages (MΦ) (CD163/CD169/CD206), myeloid dendritic cells (mDCs) (CD11c), and plasmacytoid dendritic cells (pDCs) (CD123). (C) Detailed view on the T cell zone, mainly composed of CD4 + helper T cells (T h ) and CD8 + cytotoxic T cells (T c ), mDCs (CD11c), and PCs (CD38/CD138). (D) Detailed view on the GC-mantle zone border, showing different B cells (CD11b, CD21, CD22), mDCs (CD11c), and PCs (CD38/CD138). (E) Cell annotations of three different tonsil samples plus respective bar graphs of gated cell populations, comparing the cell content between the three tonsil samples. Depicted markers and annotated cell types as indicated by the color code. ROI sizes: 976 x 640 µm, zoomed-in subregions in (C, D) : 334 µm x 219 µm. Scale bar: 100 µm.

Article Snippet: CD206 , DCN228 , 50 , 130-123-671 , FITC (PE) , Miltenyi Biotec.

Techniques: Staining, Clinical Proteomics

Journal: Heliyon

Article Title: Embelin attenuates lipopolysaccharide-induced acute kidney injury through the inhibition of M1 macrophage activation and NF-κB signaling in mice

doi: 10.1016/j.heliyon.2023.e14006

Figure Lengend Snippet: Embelin inhibits the activation of M1 macrophages in the LPS mouse model. A) M1 marker (iNOS) and M2 markers (CD163 and CD206) were detected using IHC. B) KIM-1 (purple), CD206 (red), iNOS (green), and DAPI (blue) are illustrated through multiplex immunofluorescence. All fields were captured using confocal microscope using a 60 × oil immersion lens. The images shown are representative of three independent experiments. The scale bars shown are 20 μm. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The primary antibodies included anti-KIM-1 (1:100, #NBP2-43761, Novus, USA), CD206 (1:200, #87887, Cell Signaling Technology), and iNOS (1:100, ab115819, Abcam).

Techniques: Activation Assay, Marker, Multiplex Assay, Immunofluorescence, Microscopy

Journal: iScience

Article Title: Aurkb deficiency disrupts microglial development, homeostasis and hinders remyelination following cuprizone-induced demyelination

doi: 10.1016/j.isci.2026.114718

Figure Lengend Snippet: Aurkb loss transiently elevates CD68 in homeostatic microglia but compromises its upregulation in an LPS-induced inflammation model (A–D) Adult Aurkb fl/fl and Cx3cr1 CreERT2/+ Aurkb fl/fl littermates (8-week-old) were i.p. injected with TAM for 5 consecutive days, followed by tissue collection at 1-month and 3-month post TAM induction. Representative immunofluorescence and quantification of CD68 in microglia at (A and B) 1-month and (C and D) 3-month post-TAM induction ( n = 5 mice per genotype per time point, Scale bars: 50 μm). (E–H) Neonatal Aurkb fl/fl and Cx3cr1 CreERT2/+ Aurkb fl/fl littermates were i.p. injected with TAM for 3 consecutive days at P1-P3, followed by tissue collection at P13. Representative immunofluorescence and quantification of (E and F) CD68 and (G and H) CD206 in microglia at P13 ( n = 6 mice per genotype, Scale bars: 50 μm). (I and J) Representative immunofluorescence and quantification of CD68 in microglia from adult Aurkb fl/fl and Cx3cr1 Cre/+ Aurkb fl/fl littermates ( n = 5 mice per genotype, Scale bars: 50 μm). The representative immunofluorescence image of the Cx3cr1 Cre/+ Aurkb fl/fl group is shared in C. (K and L) Adult Aurkb fl/fl and Cx3cr1 Cre/+ Aurkb fl/fl littermates were i.p. injected with LPS (1 mg/kg) and sacrificed at 48 h post LPS administration ( n = 5 mice per genotype, Scale bars: 50 μm). Data are presented as the mean ± SD. Two-tailed unpaired t-tests in (B, D, J, and I); two-way ANOVA with Bonferroni multiple comparisons test in (F, H); ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. ns, not significant compared with the Aurkb fl/fl group. See also and .

Article Snippet: The following primary antibodies were used: rabbit anti-Iba-1 antibody (1:500, Wako, Cat: 019–19741), mouse anti-Iba-1 antibody (1:400, Abcam, Cat: ab283319), rat anti-CD206 antibody (1:200, Bio-Rad, Cat: MCA2235), rat anti-BrdU antibody (1:400, Abcam, Cat: ab6326), mouse anti-phospho-Histone H3 (Ser10) antibody (1:200, Cell Signaling Technology, Cat: 9706S), mouse anti-APC (1:100, CC-1, Merck, Cat: OP80), rat anti-myelin basic protein (Mbp) monoclonal antibody (1:500, Abcam, Cat: ab7349), rabbit anti-degraded myelin basic protein (dMbp) antibody (1:2000, Millipore Sigma, Cat: AB5864), rabbit anti-Olig2 antibody (1:500, Proteintech, Cat: 13999-1-AP) and rat anti-CD68 antibody (1:500, Abcam, Cat: ab53444).

Techniques: Injection, Immunofluorescence, Two Tailed Test

Journal: PeerJ

Article Title: Cirsilineol improves anesthesia/surgery-induced postoperative cognitive dysfunction through attenuating oxidative stress and modulating microglia M1/M2 polarization

doi: 10.7717/peerj.18507

Figure Lengend Snippet: Sequences of primers.

Article Snippet: Following washing with PBS, the brain tissues and BV-2 cells were incubated with the blocking buffer at 4 °C for 1 h and the primary antibodies against Iba-1 (019-19741; 1:1,000, Wako Fujifilm, Osaka, Japan), CD86 (942-RBM-4-P1; 1:1,000, Thermo Fisher Scientific) and CD206 (CL594-60143; 1:100, Proteintech, Wuhan, China) at 4 °C overnight.

Techniques: Sequencing

Journal: PeerJ

Article Title: Cirsilineol improves anesthesia/surgery-induced postoperative cognitive dysfunction through attenuating oxidative stress and modulating microglia M1/M2 polarization

doi: 10.7717/peerj.18507

Figure Lengend Snippet: (A and B) Iba-1 + cells in each group of mice based on the quantification on the fluorescence intensity in immunofluorescence assay. (C and D) The calculated levels of microglia polarization indicators Cd86 (M1) and Cd206 (M2). (E and F) The gauged levels of SOD and MDA in plasm of mice in each group. Results were expressed as mean ± standard deviation. ns represents non-significant; P > 0.05; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Following washing with PBS, the brain tissues and BV-2 cells were incubated with the blocking buffer at 4 °C for 1 h and the primary antibodies against Iba-1 (019-19741; 1:1,000, Wako Fujifilm, Osaka, Japan), CD86 (942-RBM-4-P1; 1:1,000, Thermo Fisher Scientific) and CD206 (CL594-60143; 1:100, Proteintech, Wuhan, China) at 4 °C overnight.

Techniques: Fluorescence, Immunofluorescence, Standard Deviation

Journal: PeerJ

Article Title: Cirsilineol improves anesthesia/surgery-induced postoperative cognitive dysfunction through attenuating oxidative stress and modulating microglia M1/M2 polarization

doi: 10.7717/peerj.18507

Figure Lengend Snippet: (A–D) Quantified CD86 MFI (A, C) and CD206 MFI (B, D) in each group based on immunofluorescence assay. (E, F) The gauged levels of SOD and MDA in the supernatant of cell culture medium in each group. Results were expressed as mean ± standard deviation. ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Following washing with PBS, the brain tissues and BV-2 cells were incubated with the blocking buffer at 4 °C for 1 h and the primary antibodies against Iba-1 (019-19741; 1:1,000, Wako Fujifilm, Osaka, Japan), CD86 (942-RBM-4-P1; 1:1,000, Thermo Fisher Scientific) and CD206 (CL594-60143; 1:100, Proteintech, Wuhan, China) at 4 °C overnight.

Techniques: Immunofluorescence, Cell Culture, Standard Deviation