Journal: Journal of medical virology

Article Title: SIAH1 modulates antiviral immune responses by targeting deubiquitinase USP19.

doi: 10.1002/jmv.29523

Figure Lengend Snippet: FIGURE 1 SIAH1 positively regulates the innate antiviral response in immune cells. (A−H) Human CD14+ monocyte derived macrophages (hCD14+ MDM) or mouse bone marrow derived macrophages (mBMDM) cells were transfected with control‐siRNA or SIAH1‐siRNA and were infected with (A, B, E, F) PR8‐GFP, and (C, D, G, H) HSV‐GFP. Virus titer were taken at 12 and 24 hpi (A, C, E, G), and secreted levels of IFN‐β and IL‐6 levels were measured by specific ELISA (B, D, F, H). (I−L) RAW264.7 cells were transfected with control‐siRNA or SIAH1‐siRNA and were infected with (I, J) VSV‐GFP, and (K, L) HSV‐GFP. GFP expression, GFP absorbance, and virus titer were taken at 12 and 24 hpi (I, K), and secreted levels of IFN‐β and IL‐6 levels were measured by specific ELISA (J, L). (M, N) hCD14+ MDM (M) or RAW264.7 (N) cells were transfected with control‐siRNA or SIAH1‐siRNA and were infected with PR8‐GFP or VSV‐GFP. At 0 and 24 hpi, cells were harvested and quantitative real‐time PCR was performed to analyze the mRNA expression of antiviral genes. (O) RAW264.7 cells transfected with control‐ siRNA or SIAH1‐siRNA and were infected with PR8‐GFP and cells were harvested at the given time point after the infection. Total and phosphorylated TBK1, IRF3, and STAT1 were checked by immunoblotting. β‐actin was used as the loading control. Data information: *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 (two‐tailed Student's t‐test). Data are representative of three independent experiments, each with similar results, and expressed as the mean ± SD of two biological replicates.

Article Snippet: Primary Peripheral Blood CD14+ Monocytes, Normal, Human (ATCC PCS‐800‐010) were cultured in Alpha MEM (Welgene) supplemented with 10% FBS and 500 U/mL of human GM‐CSF (enzynomics).

Techniques: Derivative Assay, Transfection, Control, Infection, Virus, Enzyme-linked Immunosorbent Assay, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Two Tailed Test

Journal: Genome biology

Article Title: Natural display of nuclear-encoded RNA on the cell surface and its impact on cell interaction.

doi: 10.1186/s13059-020-02145-6

Figure Lengend Snippet: Fig. 2 Imaging flow cytometry (IFC) analysis of maxRNAs co-localization with immune cell surface markers in human PBMCs. a A schematic view of the extracellular hybridization of the fluorophore (red dot) labeled probes (isFISH probes) to a hypothetical maxRNA. b, c IFC images of two representative PBMC cells (rows) in 5 channels (columns), including DAPI, CD19, CD3ε, CD14, and maxRNA probes, as well as merged images of all channels (Composite). d, e The proportions of PBMCs that exhibit IFC signals (y-axis) when assayed with maxRNA probes and 3 types of controls (dArt4 probe, 6-mer library, and fluorophore only). Each box plot was derived from 4 independent biological replicates. d Total IFC positive cells PBMCs for each probe treatment. e CD14+, CD3ε+, CD19+, and CD3ε−CD14−CD19−PBMCs are separately plotted

Article Snippet: CD14+ monocytes were isolated directly from human whole blood samples using StraightFrom LRSC CD14 MicroBead Kit (Miltenyi Biotec, cat. #130-117-026), following the manufacturer’s protocol.

Techniques: Imaging, Flow Cytometry, Hybridization, Labeling, Derivative Assay

Journal: bioRxiv

Article Title: Responders vs. non-responders to mesenchymal stromal cells in knee osteoarthritis patients: mechanistic correlates of donor cell attributes and patient features

doi: 10.1101/2025.07.17.665028

Figure Lengend Snippet: A) Higher levels of resistin significantly positively correlated to delta values in KOOS Sports/Rec scores at 24 months. Higher levels of TIMP1 and CCL2 significantly correlated inversely to the percentage change and/or delta in KOOS Sports/Rec scores at 24 months relative to baseline. B) Higher levels of HGF and VEGFA in synovial fluid significantly correlated inversely to delta values and/or percentage change in KOOS Pain scores at 12 months relative to baseline. C) Higher levels of sCD14 and CCL2 significantly correlate inversely to the percentage change and/or delta in KOOS Symptom and Sports/Rec scores at 3 months relative to baseline. Delta and percent change in KOOS values were made relative to baseline and calculated such that positive values represent improvement. N=8-9 patients. Spearman’s correlation. Sports/Rec: function in sport and recreation; MO: month; KOOS: knee injury and osteoarthritis outcome score; TIMP1: tissue inhibitor of metalloproteinases 1; CCL2: chemokine C-C motif ligand 2; HGF: hepatocyte growth factor; sCD14: soluble CD14; VEGFA: vascular endothelial growth factor A.

Article Snippet: Indirect co-culture of unmatched human CD14 + peripheral blood-derived monocytes (STEMCELL Technologies, Vancouver, Canada) with biobanked MSC(M) was performed as previously described [ ].

Techniques:

Journal: Pediatric Rheumatology Online Journal

Article Title: Surface expression and genotypes of Toll- like receptors 2 and 4 in patients with juvenile idiopathic arthritis and systemic lupus erythematosus

doi: 10.1186/1546-0096-11-9

Figure Lengend Snippet: Representative flow cytometry histograms showing (a) TLR2 and (b) TLR4 expression on CD14 + monocytes. Blue filled histograms: isotype controls and green or red filled histogram TLR expression. Mean channel fluorescence intensity (MFI) derived from fluorescence histogram was used to study the level of cell surface TLR expression. Delta MFI (dMFI) was calculated as a subtraction and recorded as the MFI of the TLR2 or TLR4 antibody minus the MFI of the isotype-matched control antibody.

Article Snippet: For surface staining 100 μl of whole blood samples were incubated with the following anti-human primary antibodies for 15 minutes in the dark at 4°C: anti-CD14 monoclonal antibody (mAb; 10 μl per 100 μl blood, FITC or PE labelled, clone 8 G3; Diaclone, Besançon, France), anti-TLR2 mAb (10 μl, FITC labelled, clone HTA 125; eBioscience, San Diego, CA) or TLR4 mAb (10 μl, PE labelled, clone HTA 125; eBioscience, San Diego, CA).

Techniques: Flow Cytometry, Expressing, Fluorescence, Derivative Assay, Control

Journal: Pediatric Rheumatology Online Journal

Article Title: Surface expression and genotypes of Toll- like receptors 2 and 4 in patients with juvenile idiopathic arthritis and systemic lupus erythematosus

doi: 10.1186/1546-0096-11-9

Figure Lengend Snippet: (a) Mean fluorescence intensity (dMFI) of TLR2-expression on CD14 + monocytes of patients with JIA, SLE and healthy controls. (b) Mean fluorescence intensity (dMFI) of TLR4-expression on CD14 + monocytes of patients with JIA, SLE and healthy controls.

Article Snippet: For surface staining 100 μl of whole blood samples were incubated with the following anti-human primary antibodies for 15 minutes in the dark at 4°C: anti-CD14 monoclonal antibody (mAb; 10 μl per 100 μl blood, FITC or PE labelled, clone 8 G3; Diaclone, Besançon, France), anti-TLR2 mAb (10 μl, FITC labelled, clone HTA 125; eBioscience, San Diego, CA) or TLR4 mAb (10 μl, PE labelled, clone HTA 125; eBioscience, San Diego, CA).

Techniques: Fluorescence, Expressing

Journal: Pediatric Rheumatology Online Journal

Article Title: Surface expression and genotypes of Toll- like receptors 2 and 4 in patients with juvenile idiopathic arthritis and systemic lupus erythematosus

doi: 10.1186/1546-0096-11-9

Figure Lengend Snippet: (a) Mean fluorescence intensity (dMFI) of TLR2-expression on CD14 + monocytes of patients in various disease phases compared to TLR2-expression on monocytes of healthy controls. (b) Mean fluorescence intensity (dMFI) of TLR4-expression on CD14 + monocytes of patients in various disease phases compared to TLR4-expression on monocytes of healthy controls.

Article Snippet: For surface staining 100 μl of whole blood samples were incubated with the following anti-human primary antibodies for 15 minutes in the dark at 4°C: anti-CD14 monoclonal antibody (mAb; 10 μl per 100 μl blood, FITC or PE labelled, clone 8 G3; Diaclone, Besançon, France), anti-TLR2 mAb (10 μl, FITC labelled, clone HTA 125; eBioscience, San Diego, CA) or TLR4 mAb (10 μl, PE labelled, clone HTA 125; eBioscience, San Diego, CA).

Techniques: Fluorescence, Expressing

Journal: Nature Communications

Article Title: In depth transcriptomic profiling defines a landscape of dysfunctional immune responses in patients with VEXAS syndrome

doi: 10.1038/s41467-025-59890-0

Figure Lengend Snippet: a Overview of the experimental workflow. A figure was created with BioRender. Mizumaki, H. (2025) [ https://BioRender.com/n37z398 ]. b Hemoglobin levels (HGB), platelet counts (PLT), white blood cell counts (WBC), neutrophil counts, monocyte counts, B cell counts, NK cell counts, and T cell counts from VEXAS patients (n = 9). Background shading shows a normal reference range for each parameter. Data are presented as mean and SD. c A Uniform Manifold Approximation and Projection (UMAP) plot of 178,168 cells from all subjects (n = 14, left). UMAP plots of 125,806 peripheral blood mononuclear cells (PBMCs) derived from VEXAS patients (n = 9, upper right) and 52,362 PBMCs derived from healthy donors (n = 5, lower right). Leiden clusters based on 5’ gene expression are shown and colored by major cell types. d A violin plot showing expression distributions of selected canonical marker genes in 10 major cell types. Rows and columns represent selected marker genes and cell types, respectively. e A UMAP plot of cells with projected mutation status assignment for wild-type UBA1 (wt UBA1 ; n = 18,140 cells) and mutated UBA1 (mt UBA1 ; n = 1402 cells). f Normalized frequency of mt UBA1 cells in major cell types. Cell types with more than 100 cells were analyzed. gdT gamma delta T cells, MAIT mucosa-associated invariant T cell. Source data are provided as a Source Data file.

Article Snippet: For effector cells, CD14 + monocytes were isolated from cryopreserved PBMCs (1 × 10 7 cells) by using CD14 MicroBeads (#130-118-906, Miltenyi Biotec) following manufacturer instructions.

Techniques: Derivative Assay, Gene Expression, Expressing, Marker, Mutagenesis