murine igf-1 standard abts elisa development kit  (PeproTech)

Journal: Viruses

Article Title: Nanotechnology-Driven Strategy Against SARS-CoV-2: Pluronic F127-Based Nanomicelles with or Without Atazanavir Reduce Viral Replication in Calu-3 Cells

doi: 10.3390/v17040518

Figure Lengend Snippet: Calu-3 cells (6-well plates with 4.0 × 10 5 cells/well) were infected with SARS-CoV-2 (MOI: 2) for 1 h at 37 °C and 5% CO 2 . The supernatant was then removed, and the cells were exposed to fresh medium containing nanomicelles at 1:2 or 1:4 dilutions with or without atazanavir (ATV) at concentrations of 250 µg/mL or 125 µg/mL, respectively, and kept at 37 °C, 5% CO 2 for 48 h. ( A ) Viral titer was determined by plaque assay. ( B ) The inhibition percentage was also determined. ( C ) IL-6 and ( D ) IL-8 cytokine production was detected using a Human Standard ABTS ELISA Development kit (Peprotech, Thermo Fisher Scientific). Data are represented as mean ± SD of 6 experiments, and graphs were created using GraphPad Prism 10.1.1 software, analyzed by one-way ANOVA followed by Tukey’s post-test ( n = 6), * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: For the quantification of IL-6 and IL-8 cytokine production, the Human Standard ABTS ELISA Development kit (Peprotech, Thermo Fisher Scientific, Waltham, MA, USA) was used, following the manufacturer’s recommendations.

Techniques: Infection, Plaque Assay, Inhibition, Enzyme-linked Immunosorbent Assay, Software

Journal: bioRxiv

Article Title: Granulocyte Derived Resistin Inhibits Monocyte Maturation and Induces Immune Suppression in CMML

doi: 10.1101/2025.03.03.640303

Figure Lengend Snippet: A: Normalised expression levels of RETN in granulocytes, CD34+ HSPCs, CD14+ monocytes and whole PBMNCs from CMML patients and healthy (young and/or old) controls , , . B. Relative expression levels of RETN in granulocytes sorted from different myeloid cancer patients . aCML: atypical CML, MPN-U: MPN-unclassifiable, CNL: Chronic Neutrophilic Leukaemia, MDS/MPN-U: MDS/MPN-unclassifiable. C. Resistin protein (ng/ml) levels in healthy (N=14) and CMML (N=27) plasma samples as determined by ELISA. D. Pearson correlation between CMML granulocyte RETN expression and matched plasma resistin concentration. E & F. Kaplan-Meier overall (E) and AML-free survival plot (F) of CMML patients stratified by plasma resistin levels (High resistin N=9; Low resistin N=18). Error bars = ± SEM; ns, no significance; ****, P<0.0001.

Article Snippet: The resistin concentration was measured in the PPP supernatant using the Human Resistin Standard ABTS ELISA Development Kit (Peprotech, 900-K235), according to manufacturer’s protocol.

Techniques: Expressing, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: bioRxiv

Article Title: Granulocyte Derived Resistin Inhibits Monocyte Maturation and Induces Immune Suppression in CMML

doi: 10.1101/2025.03.03.640303

Figure Lengend Snippet: A. Representative flow cytometry plots showing the surface marker expression of CD14 and CD16 on freshly isolated healthy monocytes (0hrs) and monocytes treated with increase in concentration of resistin or control GFP for 24 hours (N=5). B & D. Bar charts showing average percentage of CD14+ or CD14+CD16+ populations under indicated conditions. C. Representative flow cytometry plots showing the surface marker expression of CD14 and CD16 on monocytes incubated with or without TLR4 neutralising antibody (TLR4Ab) prior to exposure with resistin or GFP control for 24 hours (N=4). E. Relative expression levels of RETN in three different types of human healthy monocytes . F. Representative brightfield microscope images of healthy monocytes/macrophages morphology following 6-day differentiation of monocytes treated either with 250 ng/ml of GFP control or resistin (Left). Blue arrows = attached macrophages; red arrows = suspended monocytes; scale bar 100 µm. Quantification cell confluency after 6 days treatment of monocytes either with GFP control or resistin (right). G. Percentage of live cells in suspension cultures of monocyte differentiation, treated either with GFP control or resistin. Error bars = ± SEM; ns, no significance; *, P<0.05; **, P<0.01; ***, P<0.001.

Article Snippet: The resistin concentration was measured in the PPP supernatant using the Human Resistin Standard ABTS ELISA Development Kit (Peprotech, 900-K235), according to manufacturer’s protocol.

Techniques: Flow Cytometry, Marker, Expressing, Isolation, Concentration Assay, Control, Incubation, Microscopy, Suspension

Journal: bioRxiv

Article Title: Granulocyte Derived Resistin Inhibits Monocyte Maturation and Induces Immune Suppression in CMML

doi: 10.1101/2025.03.03.640303

Figure Lengend Snippet: A. Schematic diagram depicting transcriptome study investigating downstream regulators of resistin or LPS on healthy monocytes. Monocytes treated recombinant protein GFP protein and freshly isolated monocytes were used as controls. B. Principal component analysis of the RNA-seq data. C. GO analysis of genes induced by resistin at 6 and 24 hrs together as compared with respective time point controls. Top ten significant pathways are shown. D & E. GSEA plots showing relative enrichment of indicated gene sets in different comparisons as listed. F. Heatmap showing expression of top30 DEGs across different comparisons.

Article Snippet: The resistin concentration was measured in the PPP supernatant using the Human Resistin Standard ABTS ELISA Development Kit (Peprotech, 900-K235), according to manufacturer’s protocol.

Techniques: Recombinant, Isolation, RNA Sequencing, Expressing

Journal: bioRxiv

Article Title: Granulocyte Derived Resistin Inhibits Monocyte Maturation and Induces Immune Suppression in CMML

doi: 10.1101/2025.03.03.640303

Figure Lengend Snippet: A. Venn diagram showing overlap of genes upregulated following 6 and 24 hrs treatment with either resistin or LPS in comparison with their respective time point controls. B. Venn diagram showing overlap of genes uniquely upregulated by resistin but not by LPS with genes that are upregulated in CMML monocytes . C. Relative expression levels of SEMA4A in monocytes treated with either resistin or control GFP protein. D. Relative expression levels of SEMA4A in CMML and healthy monocytes. E. Mean fluorescence intensity (MFI) of SEMA4A on healthy monocytes treated with either control GFP or increase in concentration of resistin for 24 hrs (N=4). F. Kaplan-Meier overall (left) and AML-free (right) survival plot of CMML patients stratified by SEMA4A expression levels (N=11 High; N=79 Low) G. GSEA plots showing relative enrichment of indicated gene sets in resistin (6 hrs) treated monocytes in comparison with the controls. Error bars = ± SEM; ns, no significance; *, P<0.05; **, P<0.01.

Article Snippet: The resistin concentration was measured in the PPP supernatant using the Human Resistin Standard ABTS ELISA Development Kit (Peprotech, 900-K235), according to manufacturer’s protocol.

Techniques: Comparison, Expressing, Control, Fluorescence, Concentration Assay

Journal: bioRxiv

Article Title: Granulocyte Derived Resistin Inhibits Monocyte Maturation and Induces Immune Suppression in CMML

doi: 10.1101/2025.03.03.640303

Figure Lengend Snippet: A. Ratio of Th2/Th1 cells in ex vivo cultures of whole- or CD14 depleted-PBMNCs treated with either GFP or resistin for 9 days (N=6). B. Th2/Th1 ratio from healthy (N=10) and CMML (N=8) peripheral blood (PB). C. Ratio of Th2/Th1 cells in PB of CMML patients stratified based plasma resistin low (N=4) and high levels (N=4). D. Percentage of Treg cells in healthy (N=8) vs CMML PB (N=13). E. Percentage of Treg cells in PB of CMML patients stratified based on resistin plasma concentration low (N=6) and high (N=7) against healthy controls. F. Schematic diagram of the normal monocytic maturation pathway and immune regulation versus dysregulated monocyte maturation and immune suppression found in CMML, caused by resistin that is released by immature granulocytes. Error bars = ± SEM; ns, no significance; **, P<0.01.

Article Snippet: The resistin concentration was measured in the PPP supernatant using the Human Resistin Standard ABTS ELISA Development Kit (Peprotech, 900-K235), according to manufacturer’s protocol.

Techniques: Ex Vivo, Clinical Proteomics, Concentration Assay