Journal: Clinical and Experimental Pharmacology & Physiology

Article Title: Mechanisms of IL‐17A Neutralisation in Alleviating Renal Fibrosis and Inflammation in Spontaneously Hypertensive Rats

doi: 10.1111/1440-1681.70116

Figure Lengend Snippet: Effects of IL‐17A neutralisation on macrophage polarisation in SHR renal tissues. (A) Representative IHC staining of M1 macrophage markers (iNOS and CD86) with quantitative analysis of their positive areas. (B) Representative IHC staining of M2 macrophage markers (Arg‐1 and CD163) with quantitative analysis of their positive areas. (C) Representative immunoblots and relative expression levels of iNOS, CD86, Arg‐1, and CD163 proteins. (D) Proportion of CD86 + and CD163 + cells among CD68 + macrophages. (E) mRNA expression levels of iNOS, CD86, Arg‐1, and CD163. Data are presented as mean ± SD ( n = 6).

Article Snippet: Sections were then incubated overnight at 4°C with primary antibodies against: E‐cadherin (Boster, China), Collagen III (Boster, China), inducible nitric oxide synthase (iNOS) (Boster, China), CD86 (Boster, China), arginase‐1 (Arg‐1) (Boster, China), CD163 (Boster, China), α‐smooth muscle actin (α‐SMA) (Cell Signalling Technology, USA).

Techniques: Immunohistochemistry, Western Blot, Expressing

Journal: Journal of neurovirology

Article Title: Detection of disease-associated microglia among various microglia phenotypes induced by West Nile virus infection in mice.

doi: 10.1007/s13365-023-01161-z

Figure Lengend Snippet: Fig. 2 Variety of microglia phenotypes in brains inoculated with WNV. a–d Sections from the brains of control or WNV-infected mice at 5 dpi were immunostained with antibodies to TMEM119, ARG1, SIGLEC-H, and CD11c, respectively. The black arrowheads indicate CD11c-positive cells. The black arrows indicate non-specific reac- tions. Scale bar: 50 µm

Article Snippet: The following primary antibodies were used: anti-WNV E-protein mouse monoclonal antibody (1:100; Merck Millipore, Burlington, MA, USA), anti-Iba1 rabbit polyclonal antibody (1:1000; Wako Chemicals), antiP2RY12 rabbit polyclonal antibody (1:1000; Atlas Antibodies, Bromma, Sweden), anti-TMEM119 rabbit monoclonal antibody (1:1000; Abcam, Cambridge, UK), anti-SIGLEC-H rat monoclonal antibody (1:100; Abcam), anti-ARG1 rabbit polyclonal antibody (1:2500; Atlas Antibodies), and antiCD11c rabbit polyclonal antibody (1:100; Proteintech, Rosemont, IL, USA).

Techniques: Control, Infection

Journal: Molecular cell

Article Title: Deciphering functional tumor-immune crosstalk through highly multiplexed imaging and deep visual proteomics.

doi: 10.1016/j.molcel.2024.12.023

Figure Lengend Snippet: Figure 3. T cell subsets have distinctive spatial proteome signatures in colorectal tumor microenvironment (A and B) Ranking of protein quantified in cytotoxic T cells (CTLs) (A) and helper T cells (TH) (B), ranked by median transformed intensity values detected by mass spectrometry, with cell-type-specific markers highlighted. (C and D) Volcano plots illustrating proteomic comparisons of CTLs (C) and TH (D) between the lamina propria and tumor epithelial regions, with T cell-activity- related markers highlighted. Significantly enriched proteins are denoted as yellow or green dots (two-sided t test, false discovery rate [FDR] < 0.05, S0 = 0.1, n = 3). (E and F) Gene Ontology (GO) term analysis of significantly altered proteins in CTLs (E) and TH (F) residing in the lamina propria and tumor epithelial regions.

Article Snippet: For CD4 + ARG1 staining, the sections were incubated overnight at 4 C with primary antibodies (anti-CD4-FITC, 1:100, R&D FAB8165G; anti-ARG1-PE, 1:200,Miltenyi Biotec 130-127-839) in a humidified chamber.

Techniques: Transformation Assay, Mass Spectrometry, Activity Assay

Journal: Molecular cell

Article Title: Deciphering functional tumor-immune crosstalk through highly multiplexed imaging and deep visual proteomics.

doi: 10.1016/j.molcel.2024.12.023

Figure Lengend Snippet: Figure 5. Cytotoxic T cells (CTLs) exhibit close proximity to tumors with a suppressive impact on tumor proliferation (A) Density gradient in the background indicates distance to tumor islands, with brighter yellow representing greater distance from tumor cells. Left panel only has the gradient whereas the middle, zoomed panel additionally depicts yellow masks of the CTLs and the right panel red dots of helper T cells (TH), respectively. (B) Representative images showcasing markers for T cell phenotyping. (C) T cell gating based on the expression of CD3, CD8, CD45RA, CD45RO, and PD1 to analyze the corresponding T cell subpopulations. (D) Representative images showing CD3- and CD8-positive CTLs in tumor parenchyma (TIL-CTLs) with absent CD45RA and CD45RO expression. (E) Distance heatmap depicting proximity and interactions between tumor cells and T and B lymphocyte subtypes, with lower values indicating higher proximity as in Figure 4E. (F) Histogram of Ki67/Ki67+ tumor cell number as a function of distance from RARO CTLs. Data represent three technical replicates. Statistical significance was determined using one-way ANOVA. ***p < 0.001, **p < 0.01. (G) Representative image displaying masks of tumor cells at varying proximities to RARO TIL-CTLs.

Article Snippet: For CD4 + ARG1 staining, the sections were incubated overnight at 4 C with primary antibodies (anti-CD4-FITC, 1:100, R&D FAB8165G; anti-ARG1-PE, 1:200,Miltenyi Biotec 130-127-839) in a humidified chamber.

Techniques: Expressing

Journal: Frontiers in Pharmacology

Article Title: Angelicin Alleviates Post-Trauma Osteoarthritis Progression by Regulating Macrophage Polarization via STAT3 Signaling Pathway

doi: 10.3389/fphar.2021.669213

Figure Lengend Snippet: Angelicin protectsM1 polarization and M2 macrophage polarization in vitro . (A) Schematic diagram of bone marrow macrophages (BMMs) isolation and macrophage M1/M2 polarization and repolarization strategy. (B) Representative morphology images in each group of three independent experiments. Bar represents 10 μm. (C, D) Immunostaining of enzyme-inducible nitric oxide synthase (iNOS) and CD206 (red) in isolated BMMs in different groups with quantification of iNOS + and CD206 + cell numbers. (E, F) Western blot assay and qRT-PCR assay of Arg-1, iNOS and β-actin in BMMs. (G) qRT-PCR assay of IL-1β and TNF-α expression in BMMs. (H) ELISA for IL-1β and TNF-α concentration from cellular supernatant. The data in figures represents the mean ± SD. Significant differences are indicated as ** p < 0.01. Scale bar represents 30 μm. DAPI, 4′,6-diamidino-2-phenylindole; IL, interleukin; LPS/IFN-γ, lipopolysaccharide/interferon-γ; ANG, angelicin.

Article Snippet: Anti-CD9, anti-gp130, anti-STAT3, anti-phospho-STAT3 (Ser727), anti-CD206, anti-iNOS, anti-Arg-1, anti-β-actin antibodies were purchased from Bioworld Technology (St. Louis Park, MN, United States).

Techniques: In Vitro, Isolation, Immunostaining, Western Blot, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: Frontiers in Pharmacology

Article Title: Angelicin Alleviates Post-Trauma Osteoarthritis Progression by Regulating Macrophage Polarization via STAT3 Signaling Pathway

doi: 10.3389/fphar.2021.669213

Figure Lengend Snippet: Angelicin could regulate M1/M2 polarization via STAT3 signal pathway. (A) Western blot analysis of CD9, gp130, STAT3, p-STAT3, and β-actin in BMMs treated with LPS, IL-4/IL-13, LPS + Angelicin, LPS + IL-4/IL-13 + Angelicin or LPS + IL-4/IL-13. (B) Western blot analysis of iNOS, Arg-1, STAT3, p-STAT3, and β-actin in BMMs treated with LPS, LPS + Angelicin, LPS + IL-4/IL-13 + Angelicin + AG490. (C) Western blot analysis of iNOS, Arg-1, STAT3, p-STAT3, and β-actin in M2 macrophages treated with LPS, LPS + Angelicin, LPS + IL-4/IL-13 + Angelicin + AG490. (D, E) Flow cytometry analysis of iNOS and CD206 expression of BMMs. (F) qRT-PCR assay of IL-1β and TNF-α expression in BMMs. (G) ELISA for IL-1β and TNF-α concentration from cellular supernatant.

Article Snippet: Anti-CD9, anti-gp130, anti-STAT3, anti-phospho-STAT3 (Ser727), anti-CD206, anti-iNOS, anti-Arg-1, anti-β-actin antibodies were purchased from Bioworld Technology (St. Louis Park, MN, United States).

Techniques: Western Blot, Flow Cytometry, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: Cancers

Article Title: Targeting BRF2 in Cancer Using Repurposed Drugs

doi: 10.3390/cancers13153778

Figure Lengend Snippet: List of antibodies used in this paper.

Article Snippet: Monoclonal Mouse anti-RAD51 (WB-1:1000) , GenTex Inc., USA.

Techniques: