Journal: bioRxiv

Article Title: A bacterial artificial chromosome (BAC)-vectored noninfectious replicon of SARS-CoV-2

doi: 10.1101/2020.09.11.294330

Figure Lengend Snippet: a Huh7 cells were treated with remdesivir as the indicated concentration. Four hours later, cells were co-transfected with replicon RNA (WT or SAA) and N mRNA. The luciferase activity in the supernatants was measured at the time points indicated. b Huh7 cells were co-transfected with replicon RNA (WT or SAA) and N mRNA. Eight hours later, medium was changed with remdesivir as the indicated concentration. The luciferase activity in the supernatants was measured at the time points indicated. c Huh7 cells were treated with remdesivir (10nM), IFN-α(100 U/ml), daclatasvir(1 μM), sofosbuvir(10 μM), 2CMC(50 μM). Four hours later, cells were co-transfected with replicon RNA (WT or SAA) and N mRNA. The luciferase activity in the supernatants was measured at the time points indicated. Medium was changed at 8 hours post transfection. Data are shown as mean±SD (n=3).

Article Snippet: Remdesivir (GS-5734), Daclatasvir (S1482), Sofosbuvir (GS-7977) were purchased from Selleckchem, 2’-C-Methylcytidine (HY-10468) was purchased from MedChem Express, IFN-α (11200-2) was purchased from PBL.

Techniques: Concentration Assay, Transfection, Luciferase, Activity Assay

Journal: Frontiers in Immunology

Article Title: IL-15 Harnesses Pro-inflammatory Function of TEMRA CD8 in Kidney-Transplant Recipients

doi: 10.3389/fimmu.2017.00778

Figure Lengend Snippet: IL-15-stimulated TEMRA CD8 from Kidney-Transplant (KT) Recipients induces the activation of endothelium through IFN-γ and TNF-α secretion. (A) Expression of CD25 or CD69 by CD8 T cell subsets purified from KT recipients stimulated for 24 h with plate-bound anti-CD3 (2 µg/mL) and IL-15 (10 ng/mL). Representative flow data are shown, and bars indicate mean ± SEM of pooled data ( n = 10–12). (B) Schematic overview of the strategy to assess the inflammation of human umbilical vein endothelial cell (HUVEC) by IL-15-activated CD8 T cell subsets from KT. (C) Expression of CX3CL1 by HUVEC after 6 h of culture in the presence of medium control or selective inhibitors [anti-IFN-γ (10 µg/mL) or anti-TNF-γ (10 µg/mL) mAb] and with supernatant (dilution of supernatant of 1/6) from CD8 T cell subsets from KT stimulated overnight with plate-bound anti-CD3 (2 µg/mL) and IL-15 (10 ng/mL). Bars indicate mean ± SEM of pooled data ( n = 4–6). Mann–Whitney test (A) or Kruskal–Wallis test followed by a Dunn’s Multiple Comparison Test (C) was performed (* p < 0.05, ** p < 0.01, **** p < 0.0001).

Article Snippet: Antibodies against the following proteins were purchased from BD Biosciences: CD3 (HIT3a and UCHT1), CD69 (SK7), CD25 (M-A251), CD28 (CD28.2), pSTAT5 pY694 (clone 47), p38MAPK pT180/pY182 (38/p38), TNFα (Mab11), IFN-γ (B27), and Annexin V. Antibodies against the following proteins were purchased from Miltenyi Biotech: CD8 (BW135/80), CD45RA (T6D11), CCR7 (REA108), pAKT pS473 (REA359), and pERK1/2 pT202/pY204 (REA152).

Techniques: Activation Assay, Expressing, Purification, Control, MANN-WHITNEY, Comparison

Journal: Science Progress

Article Title: NLRX1 can counteract innate immune response induced by an external stimulus favoring HBV infection by competitive inhibition of MAVS-RLRs signaling in HepG2-NTCP cells

doi: 10.1177/00368504211058036

Figure Lengend Snippet: The effect of NLRX1 on HepG2-NTCP cells after HBV serum treatment. (A) RT-qPCR was used to measure the mRNA expression of NLRX1 in HepG2-NTCP cells after treatment with or without HBV serum for 48 h. (B) Western blotting was used to determine the expression of NLRX1 in HepG2-NTCP cells after treatment with or without HBV serum for 48 h. The levels of IFN-α, IFN-β, and IL-6 were determined by RT-qPCR (C), and ELISA (D) assays in HepG2-NTCP cells after treatment with or without HBV serum for 48 h. All experiments were independently repeated for three times. Mann–Whitney U test was applied for the comparison of two sets of data.

Article Snippet: A human IFN-α ELISA kit (Cusabio, Wuhan, China, #CSB-E08636 h), a human IFN-β ELISA kit (Cusabio, CSB-E09889 h), and a human IL-6 ELISA kit (Cusabio, #CSB-E04638 h) were used to detect the levels of IFN-α, IFN-β, and IL-6 in the supernatant of HBV serum-treated HepG2-NTCP cells, according to the supplied instructions.

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Comparison

Journal: Science Progress

Article Title: NLRX1 can counteract innate immune response induced by an external stimulus favoring HBV infection by competitive inhibition of MAVS-RLRs signaling in HepG2-NTCP cells

doi: 10.1177/00368504211058036

Figure Lengend Snippet: Analysis of the antiviral capability of HepG2-NTCP cells after interference or overexpression of NLRX1. The HepG2-NTCP cells were transfected with NLRX1-overexpressed plasmid or NLRX1 siRNA for 48 h, and then were treated with HBV serum for another 48 h prior further analysis. The level of NLRX1 was obtained using RT-qPCR (A) and western blotting (B). (C) Expression of HBV DNA and HBV cccDNA were detected by PCR. (D) Western blotting was conducted to examine the protein expressions of HBsAg and HBcAg. (E) HBcAg and HBsAg particles were observed by IHC staining (hematoxylin, silver-gray particles, magnification 400 × ). The levels of IFN-α, IFN-β, and IL-6 were determined using RT-qPCR (F) and ELISA (G). All experiments were independently repeated for three times. One-way ANOVA was applied for the comparison of three sets of data.

Article Snippet: A human IFN-α ELISA kit (Cusabio, Wuhan, China, #CSB-E08636 h), a human IFN-β ELISA kit (Cusabio, CSB-E09889 h), and a human IL-6 ELISA kit (Cusabio, #CSB-E04638 h) were used to detect the levels of IFN-α, IFN-β, and IL-6 in the supernatant of HBV serum-treated HepG2-NTCP cells, according to the supplied instructions.

Techniques: Over Expression, Transfection, Plasmid Preparation, Quantitative RT-PCR, Western Blot, Expressing, Immunohistochemistry, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Veterinary Research

Article Title: Cross-species transmission and histopathological variation in specific-pathogen-free minipigs infected with different hepatitis E virus strains

doi: 10.1186/s13567-024-01337-3

Figure Lengend Snippet: Time course of serological indicators in minipigs infected with HEV from different species . All experiments were performed using plasma samples. A ALT levels. The normal range was 31–58 IU/L. B AST levels. The normal range was 32–84 IU/L. C Lipase levels. The normal range was < 100 IU/L. D IFN-α levels, E IFN-γ levels, and F seroconversion. The cut-off value was calculated as 0.5 plus the mean absorbance of the non-reactive control. * p < 0.05, *** p < 0.001

Article Snippet: To demonstrate the plasma levels of interferon-α (IFN-α) and interferon-γ (IFN-γ) and seroconversion, ELISA was performed using a Pig Interferon α, IFN-α ELISA Kit (CSB-E07328p; Cusabio Technology, Houston, TX, USA), a Porcine IFN-gamma ELISA Kit (ES9RB; Thermo Fisher Scientific, Waltham, MA, USA) and an HEV ELISA 4.0v (veterinary) (63,541–096; MP Biomedicals, Irvine, CA, USA).

Techniques: Infection, Clinical Proteomics, Control

Journal: bioRxiv

Article Title: Pan-cancer tumor classification by a holistic tumor microenvironment atlas

doi: 10.64898/2025.12.27.696641

Figure Lengend Snippet: a, GO enrichment analysis on genes differentially expressed in CXCL9 + and IFIT1 + TAMs. Red and blue represent pathways enriched in CXCL9 + and IFIT1 + TAMs, respectively. P values are adjusted by BH in the hypergeometric test. b, Dot plot showing the expression of representative signature genes of CXCL9 + and IFIT1 + TAMs. The dot size represents the proportion of expressing cells. The color indicates the average level of gene expression. c, Quantification of the mRNA expression of CXCL9 + macrophage-specific gene ( CXCL9 ) and IFIT1 + macrophage-specific genes ( IFIT1 ) in monocyte-derived macrophages, following a 24h stimulation with IFNA1, IFNA13, IFNA14, IFNB1, IFNG or control. P values are calculated using the two-sided t-test, where * indicates P < 0.05, ** indicates P < 0.01, and *** indicates P < 0.001. d, ATAC-seq tracks showing the chromatin accessibility in the CXCL9 and IFIT1 loci for macrophage subsets in TNBC tumors. e, Scatter plots showing the spatial distribution pattern of CD68 + CD163 + CXCL9 + cells and CD68 + CD163 + IFIT1 + cells in representative Xenium samples. Each dot represents an individual cell, and color represents the cell identity. The right bar plots showing the proportion of IFIT1 + or CXCL9 + macrophages across Xenium samples. f, Scatter plots showing effect sizes of the cancer cell state comparisons between c68-CXCL9 high versus c68-CXCL9 low and c69-IFIT1 high versus c69-IFIT1 low tumors. Left: recurrent cancer gene programs (Barkley et al.); right: HALLMARK pathways (MsigDB). Dots represent cancer cell states, and colors represent statistical significance categories. Effect sizes are calculated as Hedge’s g values derived from Student’s t-test, and P values are adjusted by the BH-method. Dots with a BH-adjusted P value < 0.05 and an absolute effect size >0.2 are highlighted.

Article Snippet: Recombinant proteins IFNA1 (Novoprotein, CC75), IFNA13 (MedChemExpress, HY-P72796), IFNA14 (MedChemExpress, HY-P72243), IFNB1 (PeproTech, 300-02BC), and IFNG (PeproTech, 300-02) were then added to the medium at a final concentration of 20 ng/mL.

Techniques: Expressing, Gene Expression, Derivative Assay, Control

Journal: Poultry Science

Article Title: New insights into the spleen injury by mitochondrial dysfunction of chicken under polystyrene microplastics stress

doi: 10.1016/j.psj.2024.103674

Figure Lengend Snippet: The primers involved in this study.

Article Snippet: GPX4 , 1:500 , Boster Biotechnology, China , A02059-1.

Techniques:

Journal: Poultry Science

Article Title: New insights into the spleen injury by mitochondrial dysfunction of chicken under polystyrene microplastics stress

doi: 10.1016/j.psj.2024.103674

Figure Lengend Snippet: The antibodies used in this study.

Article Snippet: GPX4 , 1:500 , Boster Biotechnology, China , A02059-1.

Techniques:

Journal: Poultry Science

Article Title: New insights into the spleen injury by mitochondrial dysfunction of chicken under polystyrene microplastics stress

doi: 10.1016/j.psj.2024.103674

Figure Lengend Snippet: MPs induced ferroptosis in spleen. (A) Western Blot results in ferroptosis-related proteins. (B) Quantitative analysis of GPX4, FTH1, and SLC7A11 protein expression. (C) Ferroptosis-related factors of mRNA expression changes. Data were expressed as mean ± SD, n = 3. *, **, ***, denotes: p < 0.05, 0.01, and 0.001, respectively.

Article Snippet: GPX4 , 1:500 , Boster Biotechnology, China , A02059-1.

Techniques: Western Blot, Expressing