Journal: Virology Journal

Article Title: Herpes simplex virus type 2 infection increases human immunodeficiency virus type 1 entry into human primary macrophages

doi: 10.1186/1743-422X-8-166

Figure Lengend Snippet: HSV-2 ability to infect human monocyte cell line U937 relies on their differentiation level . A) HSV-2 replication in undifferentiated U937 cells. The U937 cell line was infected with two different MOI of HSV-2, as reported in the graph legend. Vero cells were infected as a control. HSV-2 titre in the cellular supernatants was measured by plaque assay, at different times post-infection (p.i.). B) Effect of TPA treatment of U937 cells on CD14 expression. The effect of TPA treatment on U937 cell differentiation state was analyzed by CD14 FACS analysis. The percentage of positive cells is reported. C) HSV-2 replication in TPA-treated U937 cells. Undifferentiated or TPA-treated U937 cells were infected with HSV-2 (MOI of 1 PFU/cell). HSV-2 titre in the cellular supernatant was measured by plaque assay. In all cases, the reported values represent the mean of four independent experiments. The error bars represent the standard deviation.

Article Snippet: Briefly, 1 × 10 6 cells were harvested and directly incubated for one hour in cold PBS containing 1:100 (v/v) of an anti-human CD14 primary antibody (Li StarFISH).

Techniques: Infection, Control, Plaque Assay, Expressing, Cell Differentiation, Standard Deviation

Journal:

Article Title: GM-CSF Regulates a PU.1-Dependent Transcriptional Program Determining the Pulmonary Response to LPS

doi: 10.1165/rcmb.2006-0174OC

Figure Lengend Snippet: GM-CSF, via PU.1, regulates mRNA levels for multiple components of the TLR-4 signaling pathway in AMs. mRNA transcripts encoding both positive and negative signaling components of the TLR-4 signaling pathway in cultured alveolar macrophages were detected by RT-PCR. Note the absence of transcripts for CD14, RP105, and IRAK-M in mAM cells.

Article Snippet: Briefly, primary antibodies included rat anti-murine CD14 (PharMingen, San Diego, CA) diluted 1:200, rabbit anti-mouse IRAK-M (Chemicon International, San Diego, CA) diluted 1:1,000, or goat anti-murine β-actin antibody (Santa Cruz, Santa Cruz, CA) diluted 1:200.

Techniques: Cell Culture, Reverse Transcription Polymerase Chain Reaction

Journal:

Article Title: GM-CSF Regulates a PU.1-Dependent Transcriptional Program Determining the Pulmonary Response to LPS

doi: 10.1165/rcmb.2006-0174OC

Figure Lengend Snippet: GM-CSF, via PU.1, regulates multiple components of TLR-4 signaling pathway in AMs. (A) Evaluation of RP105 on cultured alveolar macrophage cell lines by flow cytometry. Cells were immunostained with PE-labeled, anti-RP105 (black line) or isotype control (gray shading) antibodies and evaluated by flow cytometry. (B) RP105 levels on AMs were quantified by determining the difference between the mean fluorescence of respective cell lines immunostained with anti-RP105 antibody minus the isotype control antibody. RP105 was readily detectible on MH-S and mAMPU.1+ cells and absent on mAM cells (n = 3 determinations/cell line; P < 0.001 [Kruskal-Wallis ANOVA on ranks]). (C) Evaluation of CD14 and IRAK-M on cultured alveolar macrophage cell lines by Western blot analysis.

Article Snippet: Briefly, primary antibodies included rat anti-murine CD14 (PharMingen, San Diego, CA) diluted 1:200, rabbit anti-mouse IRAK-M (Chemicon International, San Diego, CA) diluted 1:1,000, or goat anti-murine β-actin antibody (Santa Cruz, Santa Cruz, CA) diluted 1:200.

Techniques: Cell Culture, Flow Cytometry, Labeling, Fluorescence, Western Blot

Journal: Cell

Article Title: SARS-CoV-2 infection triggers profibrotic macrophage responses and lung fibrosis

doi: 10.1016/j.cell.2021.11.033

Figure Lengend Snippet:

Article Snippet: CD14-PE , Miltenyi Biotec , Cat# 130-113-709, RRID: AB_2726250.

Techniques: Immunohistochemistry, Plasmid Preparation, Recombinant, Staining, Lysis, Protease Inhibitor, Mass Spectrometry, Sequencing, Modification, Bicinchoninic Acid Protein Assay, Enzyme-linked Immunosorbent Assay, Software

Journal: Hepatology (Baltimore, Md.)

Article Title: Human macrophage ferroportin biology and the basis for the ferroportin disease

doi: 10.1002/hep.29007

Figure Lengend Snippet: Liver and macrophage phenotype of FD patients. (A) Liver biopsy sections from patients with FD investigated in this study stained with Perls’ Prussian blue for iron. Pictures show the classic iron‐laden phenotype of Kupffer cells (arrows) in p.A77D, p.G80S, and pVal162del FD patients. On the contrary, microphotograph from a patient carrying the p.A69T FPN1 mutation shows iron accumulation in hepatocytes, but not in Kupffer cells. Donor and disease PBMC‐derived macrophages (B) express after 7 days the CD14 macrophage‐specific marker and (C) retain normal phagocytic activity when incubated in the presence of autofluorescent beads (see Patients and Methods). (D) Ferritin content of representative macrophage preparations. The amount of intracellular ferritin is doubled in macrophages from lack‐of‐function FD patients and halved in macrophages from patients carrying the Hepc‐resistant p.A69T mutation. Ferritin values are mean ± SEM of three different cell preparations from a representative patient as compared to cell preparations from a blood donor. * P < 0.05.

Article Snippet: The primary antibodies used were: mouse antihuman CD14 (1:20; Chemicon International, Temecula, CA); mouse anti–sodium‐potassium pump (Na,K‐ATPase [plasma membrane marker]; 1:500, Abcam, Cambridge, UK); rabbit anticalnexin (1:200 [endoplasmic reticulum marker]; Sigma‐Aldrich); rabbit anti‐golgin‐97 (1:100 [Golgi marker]; Molecular Probes, Leiden, The Netherlands )(Golgi marker); rabbit anti–early endosome antigen 1 (EEA‐1; 1:200; Abcam, Cambridge, UK), as an early endosomes marker; rat anti–lysosomal‐associated membrane protein 1 (LAMP‐1 [lysosome marker]; 1:50; Fitzgerald Industries International, Concord, MA); rabbit anti‐FPN1 (1:50 PBS; Alpha Diagnostic International, San Antonio, TX, USA); rabbit anti–human transferrin receptor 1 (TfR1; 1:50; Santa Cruz Biotechnology, Santa Cruz, CA); and rabbit anti‐Hepc (1:50 PBS; Alpha Diagnostic International).

Techniques: Staining, Mutagenesis, Derivative Assay, Marker, Activity Assay, Incubation