Journal: bioRxiv

Article Title: ARG1 could be expressed by human, but not represent for repair

doi: 10.1101/2020.07.02.183624

Figure Lengend Snippet: (A) The schematic diagram shows the time point after the pre-operation of GBR when tissues were harvested and the assays performed. (B and C) Analysis of RNA-seq data: both fpkm (B) and gene count (C) confirmed that ARG1 was not detected in the normal and inflammatory states, and its expression was low at the healing site, even when compared with other genes related to macrophages. (D) qPCR assay of ARG1 showed no significant differences between tissues from inflammatory and healing sites. Tissues were collected and digested to obtain single-cell suspensions for flow cytometry analysis. CD68and CD11b were used as markers for myeloid macrophages. iNOS and ARG1 were used as markers for M1 and M2 macrophages. (E) ARG1 expression was not detected in either group, and CD11b + CD68 + ARG1 + iNOS − macrophages were not observed using gated and t-distributed stochastic neighbor embedding (t-SNE) dimension reduction to visualize high-dimensional data. (F) Immunofluorescent staining did not reveal ARG1 in any group. Hoechst staining appears as blue in color while ARG1 appears as red. (G) Detection of surface markers by flow cytometry analysis. High expression of CD68 (CD68-FITC, Clone Y1/82A) and low expression of Arg1 were evident in the three groups. The population size of CD11b + CD68 + ARG1 − iNOS + macrophages was very small. Abbreviations: NB, normal alveolar bone tissue; IP, inflammatory periodontal tissue; H, healing (tissue harvest after guided bone regeneration surgery), and N/A, undetected.

Article Snippet: Cells were isolated by trypsinization after cultured for 1 (THP-1 monocytes to macrophages) and 7 (THP-1 monocytes derived macrophages after culturing in different circumstances) days respectively, co-incubated with antibodies against iNOS (iNOS-PE, Clone 4E5, Novus Biologicals, USA), CD68 (CD68-FITC, Clone Y1/82A, Abcam, UK), CD11b (PerCP/Cy5.5®, Clone M1/70, Abcam, UK) and ARG1 (ARG1-PE/Cyanine, Clone 14D2C43, BioLegend, USA) at 1:400 dilution in the dark for 1 h at 4°C (100μl per antibody for each sample).

Techniques: RNA Sequencing, Expressing, Flow Cytometry, Staining

Journal: bioRxiv

Article Title: ARG1 could be expressed by human, but not represent for repair

doi: 10.1101/2020.07.02.183624

Figure Lengend Snippet: (A) Schematic diagram of the cell culture process and the time points for stimulation of macrophage differentiation, seeding macrophages in the different conditions, and performing the assays. (B) Light microscopy images of macrophages cultured under different conditions on Day 7. (C) Overlay of tSNE dimensionality reduction diagrams revealed the absence of AGR1 + macrophages in the flow cytometry analysis. (D) qPCR assay of ARG1 using Gapdh as the reference gene did not detect ARG1 in the four groups. (E-G) In contrast to the large population of CD11b + CD68 + macrophages (E) and CD68 + CD11b + iNOS + ARG − (F), CD68 + CD11b + iNOS − ARG1 + macrophages were not detected (G). Abbreviations: Ctrl, RPMI supplemented with 10% FBS; LPS, RPMI supplemented with 10% FBS and 10 ng/ml LPS; IL4, RPMI supplemented with 10% FBS, 10 ng/ml IL4; DBBM, RPMI supplemented with 10% FBS and 0.03 g/well DBBM; NS, no significance; N/A, undetected. *** denotes P < 0.001.

Article Snippet: Cells were isolated by trypsinization after cultured for 1 (THP-1 monocytes to macrophages) and 7 (THP-1 monocytes derived macrophages after culturing in different circumstances) days respectively, co-incubated with antibodies against iNOS (iNOS-PE, Clone 4E5, Novus Biologicals, USA), CD68 (CD68-FITC, Clone Y1/82A, Abcam, UK), CD11b (PerCP/Cy5.5®, Clone M1/70, Abcam, UK) and ARG1 (ARG1-PE/Cyanine, Clone 14D2C43, BioLegend, USA) at 1:400 dilution in the dark for 1 h at 4°C (100μl per antibody for each sample).

Techniques: Cell Culture, Light Microscopy, Flow Cytometry

Journal: Molecular Imaging and Biology

Article Title: 124 I-Iodo-DPA-713 Positron Emission Tomography in a Hamster Model of SARS-CoV-2 Infection

doi: 10.1007/s11307-021-01638-5

Figure Lengend Snippet: Iba-1 staining and flow cytometry. a Immunohistochemistry demonstrates clusters of Iba-positive cells within pneumonic tissues. b Flow cytometry demonstrates a higher percentage of myeloid cells and CD11b + cells (macrophages, phagocytes) in males ( n = 5), than females ( n = 5). Data represented as median ± interquartile range.

Article Snippet: A single-cell suspension was stained with Zombie Aqua Fixable Viability Kit (Biolegend), resuspended in FACS buffer (1% bovine serum albumin in saline), and incubated in block buffer prior to surface staining with anti-CD11b antibody (Novus Biologicals, #NB110-89474PECY7, polyclonal) for macrophages, monocytes, and granulocytes.

Techniques: Staining, Flow Cytometry, Immunohistochemistry

Journal: iScience

Article Title: Intravenous BCG vaccination reduces SARS-CoV-2 severity and promotes extensive reprogramming of lung immune cells

doi: 10.1016/j.isci.2023.107733

Figure Lengend Snippet:

Article Snippet: The antibodies used in this study were purchased from different vendors as follows: Flow cytometry antibodies: Anti-CD3 (MCA1477PB, CD3-12): Bio Rad; Anti-CD4 (100451, GK1.5): BioLegend; Anti-CD11b (NB110-89474PECY7, Poly): Novus; Anti-RT1D (110211, 14-4-4S): BioLegend.

Techniques: Virus, Recombinant, Lysis, Red Blood Cell Lysis, Cell Viability Assay, Neutralization, Software