apc cd206  (R&D Systems)

Journal: Antioxidants

Article Title: Caffeic Acid Phenethyl Ester Suppresses Oxidative Stress and Regulates M1/M2 Microglia Polarization via Sirt6/Nrf2 Pathway to Mitigate Cognitive Impairment in Aged Mice following Anesthesia and Surgery

doi: 10.3390/antiox12030714

Figure Lengend Snippet: All primary and secondary antibodies used in this study.

Article Snippet: After being washed with DPBS, a membrane-breaking fixative solution (BD Bioscience, 554714, San Jose, CA, USA) was used to fix for 15 min. Flow cytometry antibodies PE CD86 (R & D Systems, FAB741P, Minneapolis, MN, USA) and APC CD206 (R & D Systems, FAB2535A, Minneapolis, MN, USA), Rat IgG2A PE-conjugated Antibody (R & D Systems, IC006P, Minneapolis, MN, USA) and Goat IgG APC-conjugated Antibody (R & D Systems, IC108A, Minneapolis, MN, USA) were used in accordance with the instructions.

Techniques:

Journal: Antioxidants

Article Title: Caffeic Acid Phenethyl Ester Suppresses Oxidative Stress and Regulates M1/M2 Microglia Polarization via Sirt6/Nrf2 Pathway to Mitigate Cognitive Impairment in Aged Mice following Anesthesia and Surgery

doi: 10.3390/antiox12030714

Figure Lengend Snippet: Primers used in this study.

Article Snippet: After being washed with DPBS, a membrane-breaking fixative solution (BD Bioscience, 554714, San Jose, CA, USA) was used to fix for 15 min. Flow cytometry antibodies PE CD86 (R & D Systems, FAB741P, Minneapolis, MN, USA) and APC CD206 (R & D Systems, FAB2535A, Minneapolis, MN, USA), Rat IgG2A PE-conjugated Antibody (R & D Systems, IC006P, Minneapolis, MN, USA) and Goat IgG APC-conjugated Antibody (R & D Systems, IC108A, Minneapolis, MN, USA) were used in accordance with the instructions.

Techniques:

Journal: Antioxidants

Article Title: Caffeic Acid Phenethyl Ester Suppresses Oxidative Stress and Regulates M1/M2 Microglia Polarization via Sirt6/Nrf2 Pathway to Mitigate Cognitive Impairment in Aged Mice following Anesthesia and Surgery

doi: 10.3390/antiox12030714

Figure Lengend Snippet: CAPE pretreatment promotes the switch of hippocampal microglia from the M1 to the M2 type after anesthesia and surgery in aged mice. ( A ) Representative immunofluorescence and 3D reconstitution images of Iba-1 + microglia. ( B ) The number of microglia in the CA1, CA3, and DG areas (two-way ANOVA, regions: p = 0.5884, groups: p value < 0.0001; interaction: p value = 0.6376; n = 6 per group). ( C ) RT-qPCR analysis of CD86 gene expression (one-way ANOVA, p value < 0.0001; n = 8 per group). ( D ) RT-qPCR analysis of iNOS gene expression (one-way ANOVA, p value < 0.0001; n = 8 per group). ( E ) RT-qPCR analysis of CD32 gene expression (one-way ANOVA, p value < 0.0001; n = 8 per group). ( F ) RT-qPCR analysis of IL-1β gene expression (one-way ANOVA, p value = 0.0071; n = 8 per group). ( G ) RT-qPCR analysis of TNF-α gene expression (one-way ANOVA, p value < 0.0001; n = 8 per group). ( H ) RT-qPCR analysis of CD206 gene expression (one-way ANOVA, p value = 0.0012; n = 8 per group). ( I ) RT-qPCR analysis of ARG-1 gene expression (one-way ANOVA, p value < 0.0001; n = 8 per group). ( J ) RT-qPCR analysis of TGF-β gene expression (one-way ANOVA, p value < 0.0001; n = 8 per group). ( K ) RT-qPCR analysis of IL-4 gene expression (one-way ANOVA, p value < 0.0001; n = 8 per group). ( L ) RT-qPCR analysis of IL-10 gene expression (one-way ANOVA, p value < 0.0001; n = 8 per group). (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; NS regarded as not significant).

Article Snippet: After being washed with DPBS, a membrane-breaking fixative solution (BD Bioscience, 554714, San Jose, CA, USA) was used to fix for 15 min. Flow cytometry antibodies PE CD86 (R & D Systems, FAB741P, Minneapolis, MN, USA) and APC CD206 (R & D Systems, FAB2535A, Minneapolis, MN, USA), Rat IgG2A PE-conjugated Antibody (R & D Systems, IC006P, Minneapolis, MN, USA) and Goat IgG APC-conjugated Antibody (R & D Systems, IC108A, Minneapolis, MN, USA) were used in accordance with the instructions.

Techniques: Immunofluorescence, Quantitative RT-PCR, Gene Expression

Journal: Antioxidants

Article Title: Caffeic Acid Phenethyl Ester Suppresses Oxidative Stress and Regulates M1/M2 Microglia Polarization via Sirt6/Nrf2 Pathway to Mitigate Cognitive Impairment in Aged Mice following Anesthesia and Surgery

doi: 10.3390/antiox12030714

Figure Lengend Snippet: CAPE promotes the switch of H 2 O 2 -induced BV2 cells from the M1 to the M2 type through activating Sirt6. ( A ) Representative flow cytometry analysis of BV2 cells induced by four conditions. ( B ) Quantification of the percentage of CD86 + cells after treatment with four conditions (one-way ANOVA, p value < 0.0001; n = 5 per group). ( C ) Quantification of the percentage of CD206 + cells after treatment with four conditions (one-way ANOVA, p value < 0.0001; n = 5 per group). ( D ) The ratio of CD86 + /CD206 + cells after treatment with four conditions (one-way ANOVA, p value < 0.0001; n = 5 per group). ( E ) Immunofluorescence staining of CD86 and Iba1 in BV2 cells after treatment with the four conditions. ( F ) Immunofluorescence staining of CD206 and Iba1 in BV2 cells after treatment with the four conditions. ( G ) Quantification of CD86 fluorescence intensity mean value in the four conditions (one-way ANOVA, p value < 0.0001; n = 4 per group). ( H ) Quantification of CD206 fluorescence intensity mean value in the four conditions (one-way ANOVA, p value < 0.0001; n = 4 per group). ( I ) RT-qPCR analysis of CD86 in BV2 cells under four conditions (one-way ANOVA, p value < 0.0001; n = 4 per group). ( J ) RT-qPCR analysis of iNOS in BV2 cells under four conditions (one-way ANOVA, p value = 0.0004; n = 4 per group). ( K ) RT-qPCR analysis of CD32 in BV2 cells under four conditions (one-way ANOVA, p value = 0.0002; n = 4 per group). ( L ) RT-qPCR analysis of CD206 in BV2 cells under four conditions (one-way ANOVA, p value < 0.0001; n = 4 per group). ( M ) RT-qPCR analysis of ARG-1 in BV2 cells under four conditions (one-way ANOVA, p value = 0.0001; n = 4 per group). ( N ) RT-qPCR analysis of TGF-β in BV2 cells under four conditions (one-way ANOVA, p value = 0.0002; n = 4 per group). ( O ) RT-qPCR analysis of the pro-inflammatory cytokine TNF-α in BV2 cells under four conditions (one-way ANOVA, p value = 0.0161; n = 4 per group). ( P ) RT-qPCR analysis of the anti-inflammatory cytokine IL-4 in BV2 cells under four conditions (one-way ANOVA, p value = 0.0018; n = 4 per group). Five or four independent experiments were done and for each, three replicates were plated. (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; NS regarded as not significant).

Article Snippet: After being washed with DPBS, a membrane-breaking fixative solution (BD Bioscience, 554714, San Jose, CA, USA) was used to fix for 15 min. Flow cytometry antibodies PE CD86 (R & D Systems, FAB741P, Minneapolis, MN, USA) and APC CD206 (R & D Systems, FAB2535A, Minneapolis, MN, USA), Rat IgG2A PE-conjugated Antibody (R & D Systems, IC006P, Minneapolis, MN, USA) and Goat IgG APC-conjugated Antibody (R & D Systems, IC108A, Minneapolis, MN, USA) were used in accordance with the instructions.

Techniques: Flow Cytometry, Immunofluorescence, Staining, Fluorescence, Quantitative RT-PCR

Journal: Antioxidants

Article Title: Caffeic Acid Phenethyl Ester Suppresses Oxidative Stress and Regulates M1/M2 Microglia Polarization via Sirt6/Nrf2 Pathway to Mitigate Cognitive Impairment in Aged Mice following Anesthesia and Surgery

doi: 10.3390/antiox12030714

Figure Lengend Snippet: All primary and secondary antibodies used in this study.

Article Snippet: Mouse MMR/CD206 APC-conjugated Antibody , Goat , , 1:20 , R & D Systems , FAB2535A.

Techniques:

Journal: Antioxidants

Article Title: Caffeic Acid Phenethyl Ester Suppresses Oxidative Stress and Regulates M1/M2 Microglia Polarization via Sirt6/Nrf2 Pathway to Mitigate Cognitive Impairment in Aged Mice following Anesthesia and Surgery

doi: 10.3390/antiox12030714

Figure Lengend Snippet: All primary and secondary antibodies used in this study.

Article Snippet: Mouse MMR/CD206 APC-conjugated Antibody , Goat , , 1:20 , R & D Systems , FAB2535A.

Techniques:

Journal: Science Advances

Article Title: Macrophages orchestrate the expansion of a proangiogenic perivascular niche during cancer progression

doi: 10.1126/sciadv.abg9518

Figure Lengend Snippet: ( A and B ) Box and whisker plots (A) and scatter plot (B) showing normalized mean M1- and M2-associated gene scores across the indicated TAM clusters identified using scRNA-seq. ( C and D ) Subset unique, significantly up-regulated GO terms (C) and individual genes (D) between the two subsets of protumoral TAM. ( E ) FACS-gated live [7-aminoactinomycin D–negative (7AAD − )] F4/80 hi TAMs from enzyme-dispersed MMTV-PyMT tumors separated on the basis of CD206 and MHCII expression (left) and assessed for Lyve-1 expression (right; color-shaded histograms) against that of the fluorescence minus one staining (FMO) control (open black line). ( F ) Quantification of the gated populations in (E) ( n = 4 tumors). ( G ) PCA plot of the 2000 most variable genes from the bulk-sequenced TAM populations ( n = 5 tumors), using CD206 − and MHCII − TAMs as a comparator. ( H ) Heatmaps comparing the relative expression of selected differentially expressed genes identified in the scRNA-seq (left) and bulk RNA-seq (right); population color is indicative of the populations identified in (G). ( I ) Representative image of a frozen section of MMTV-PyMT tumor showing DAPI (nuclei; blue); intravenous dextran marking vasculature (green), F4/80 (magenta), and Lyve-1 (red); and colocalizing pixels for Lyve-1 and F4/80 (white); scale bars, 25 μm. ( J to M ) Schematic for experimental approach to label pvTAMs using Dil-labeled liposomes (J). (K) Representative images of frozen sections of MMTV-PyMT tumors showing DAPI (nuclei; blue); intravenous dextran marking vasculature (green), Dil (red), and F4/80 (magenta); and Dil/F4/80 colocalizing pixels (white) (right panel alone); scale bars, 25 μm (left) and 50 μm (right). (L) Quantification of the spatial location of Dil + F4/80 + TAMs ( n = 5 mice). (M) Analysis of the surface phenotype of Dil +/− TAM from enzyme-dispersed tumors within the F4/80 + gate. Box and whisker plots; boxes show median and quartiles. Bar charts represent mean, and the dots show individual tumors and mice. **** P < 0.0001.

Article Snippet: The following antibodies against the indicated antigen were purchased from Thermo Fisher Scientific and were used at 1 μg/ml unless stated otherwise: CD3ε allophycocyanin (APC) and phycoerythrin (PE) (145-2C11), CD4 FITC (RM4-5), CD8β eFluor 450 (H35-17.2), CD11b APC–eFluor 780 (M1/70), CD11b BV510 (M1/70), CD11c APC (N418), CD16/32 (2.4G2; Tonbo Biosciences), CD19 APC (6D5; BioLegend), CD29 APC (eBioHMb1-1), CD31 eFluor 450 and PE (390), CD34 FITC and APC (RAM34), CD45 APC–eFluor 780, FITC, and peridinin chlorophyll protein (PerCP)–Cy5.5 (30-F11), CD90.2 eFluor 450 (53-2.1), CD90.1 eFluor 450 (HIS51), CD90.1 BV510 (OX-7), CD103 PE (2E7), CD206 APC (FAB2535A; Bio-Techne), F4/80 PE (BM8; BioLegend), F4/80 BV421 (BM8; BioLegend), FAP (10 μg/ml; AF3715, Bio-Techne), Ly6C PE and eFluor 450 (HK1.4), Ly6G FITC (1A8; BioLegend), Lyve-1 Alexa Fluor 488 (ALY7), MHCII PE, FITC, and eFluor 450 (M5/114.15.2), NG2 Alexa Fluor 488 (AB5320A4; Millipore), NK1.1 APC (PK136), PDGFRα APC and PerCP-Cy5.5 (APA5), PDGFRβ PE (APB5), and Ly6A/E Alexa Fluor 700 (D7).

Techniques: Whisker Assay, Expressing, Fluorescence, Staining, RNA Sequencing Assay, Labeling, Liposomes