Journal: iScience

Article Title: C9ORF72 suppresses JAK-STAT mediated inflammation

doi: 10.1016/j.isci.2023.106579

Figure Lengend Snippet:

Article Snippet: rabbit anti-pSTAT1 , R and D Systems , Cat# AF2894, RRID: AB_2198137.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Reverse Transcription, Bradford Protein Assay, CRISPR, Plasmid Preparation, Software

Journal: Journal of Virology

Article Title: Tissue-Specific Gene Expression during Productive Human Papillomavirus 16 Infection of Cervical, Foreskin, and Tonsil Epithelium

doi: 10.1128/JVI.00915-19

Figure Lengend Snippet: Expression levels of total STAT1, pSTAT1, total SMAD3, and pSMAD3 in uninfected and HPV16-infected rafts. (a) Representative blots indicating expression levels of total STAT1, pSTAT1, total SMAD3, and pSMAD3 in HPV16-infected cervical, foreskin, and tonsil rafts harvested at day 10. GAPDH was used as a loading control. (b to d) Histograms representing average ratios of protein expression levels normalized to GAPDH levels (means and SEM) in cervical (b), foreskin (c), and tonsillar (d) tissues. * indicates a P value of <0.05.

Article Snippet: GraphPad Prism was used to plot and analyze quantification data. summarizes the antibodies used for the study. table ft1 table-wrap mode="anchored" t5 TABLE 5 caption a7 Protein a Dilution Catalogue number; manufacturer KRT14 1:3,000 C-8791; Sigma KRT19 1:2,000 sc-6278; Santa Cruz KRT23 1:2,000 H00025984-M01; Abnova GPER 1:1,000 MAB5534; R&D Systems STAT1 1:2,000 9172; Cell Signaling Technology pSTAT1 1:2,000 sc-8394; Santa Cruz SMAD3 1:2,000 9513; Cell Signaling Technology pSMAD3 1:2,000 9520; Cell Signaling Technology GAPDH 1:2,000 sc-47724; Santa Cruz Open in a separate window a GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Techniques: Expressing, Infection

Journal: Journal of Virology

Article Title: Tissue-Specific Gene Expression during Productive Human Papillomavirus 16 Infection of Cervical, Foreskin, and Tonsil Epithelium

doi: 10.1128/JVI.00915-19

Figure Lengend Snippet: Antibodies used

Article Snippet: GraphPad Prism was used to plot and analyze quantification data. summarizes the antibodies used for the study. table ft1 table-wrap mode="anchored" t5 TABLE 5 caption a7 Protein a Dilution Catalogue number; manufacturer KRT14 1:3,000 C-8791; Sigma KRT19 1:2,000 sc-6278; Santa Cruz KRT23 1:2,000 H00025984-M01; Abnova GPER 1:1,000 MAB5534; R&D Systems STAT1 1:2,000 9172; Cell Signaling Technology pSTAT1 1:2,000 sc-8394; Santa Cruz SMAD3 1:2,000 9513; Cell Signaling Technology pSMAD3 1:2,000 9520; Cell Signaling Technology GAPDH 1:2,000 sc-47724; Santa Cruz Open in a separate window a GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Techniques:

Journal: Frontiers in Immunology

Article Title: Hypothermia Promotes Interleukin-22 Expression and Fine-Tunes Its Biological Activity

doi: 10.3389/fimmu.2017.00742

Figure Lengend Snippet: Effects of hypothermic preconditioning on interleukin (IL)-22 bioactivity. Epithelial (-like) IL-22-responsive human DLD1 (A) and Caco2 colon (B) carcinoma cells as well as HepG2 hepatoma cells (C) were preincubated for 6 h without additional stimulation at 30 or 37°C. Thereafter, cells maintained under the respective ambient temperatures were further kept as unstimulated control or stimulated with IL-22 (20 ng/ml). After 1 or 3 h, cells were harvested and IL-22-induced signal transducer and activator of transcription (STAT)-3 activation was determined by immunoblot analysis for pSTAT3. (A–C) Densitometric quantification of experiments ( n = 3) is depicted as raw data (means ± SD). ** P < 0.01, *** P < 0.001 compared to unstimulated control at the respective temperature and time point; # P < 0.05, ## P < 0.01, ### P < 0.001; statistical analysis for individual time points, one-way ANOVA with post hoc Bonferroni correction. (D) One representative of the three independently performed experiments (for each cell type) is shown. (E–H) DLD1 cells were preincubated for 6 h without additional stimulation at 30 or 37°C. (E) Thereafter, cells were further kept under the respective ambient temperatures as unstimulated control or were stimulated with IL-22 (20 ng/ml). After the indicated time periods, α1ACT mRNA expression was determined by real-time polymerase chain reaction (PCR) using GAPDH for normalization. Data are shown as means ± SD [1 h, n = 5 (30 and 37°C); 4 h, n = 8 (30°C) and n = 7 (37°C); 12 h, n = 8 (30 and 37°C); 24 h, n = 6 (30°C), and n = 4 (37°C)]; * P < 0.05, ** P < 0.01; statistical analysis on fold-induction at each time point, unpaired Student’s t -test. AUC, area under the curve. (F) Thereafter, cells were further kept under the respective ambient temperatures as unstimulated control or stimulated with interferon (IFN)γ (20 ng/ml). After 1 or 3 h, cells were harvested and IFNγ-induced STAT1 activation was determined by immunoblot analysis for pSTAT1. One representative of three independently performed experiments is shown. Densitometric quantification of these experiments ( n = 3) is depicted in (G) as raw data (means ± SD). ** P < 0.01, *** P < 0.001 compared to unstimulated control at the respective temperature and time point; ## P < 0.01; statistical analysis for individual time points, one-way ANOVA with post hoc Bonferroni correction. (H) Thereafter, cells were further kept under the respective ambient temperatures as unstimulated control or stimulated with IFNγ (20 ng/ml). After 4 or 12 h, IL-18 binding protein (IL-18BP) mRNA expression was determined by real-time PCR using GAPDH for normalization. Data are shown as means ± SD (4 h, n = 4; 12 h, n = 5); * P < 0.05, ** P < 0.01 compared to unstimulated control of the respective temperature and time point; # P < 0.05; statistical analysis on raw data for individual time points, one-way ANOVA with post hoc Bonferroni correction.

Article Snippet: Antibodies: total STAT3-#124H6 (mouse monoclonal antibody); total STAT1, pSTAT1-Y701-#D4A7 (both rabbit polyclonal antibodies); pSTAT3-Y705-#D3A7 (rabbit monoclonal antibody); all from Cell Signaling, Frankfurt, Germany.

Techniques: Control, Activation Assay, Western Blot, Expressing, Real-time Polymerase Chain Reaction, Binding Assay