Journal: medRxiv

Article Title: Polymorphism in IFNAR contributes to glucocorticoid response and outcome in ARDS and COVID-19

doi: 10.1101/2022.03.10.22272123

Figure Lengend Snippet: (A ) STAT1 expression in the lung after 4-day culture in the presence of IFN beta with or without hydrocortisone (HC). ( B) pSTAT1 expression in the same specimens as in A. ( C) Example photomicrographs showing higher STAT2 expression in a TT patient than in a CT patient and the effect of HC on its nuclear translocation. Most STAT2 remains in the cytoplasm of the CT patients, whereas nuclear expression is prominent in the TT patient. Indicated insets are shown in the bottom row. Arrows. ( D ) Combined results of all patients noting that two CT samples are excluded in the data as the patients were already under glucocorticoid treatment at the time of sample acquisition. Ns, not significant; *P<0.05; **P<0.01; and ***P<0.001

Article Snippet: The first stage antibodies were anti-alpha chain of the IFN alpha/beta receptor (St John’s Laboratory STJ112765) that was used 1:2000 and 1:5000 and anti-Stat1 (1:400, 9175S), anti-pStat1(1:100, 9167S) and anti-Stat2 (1:200, 72604S) all from Cell Signalling.

Techniques: Expressing, Translocation Assay

Journal: Oncotarget

Article Title: PTEN mediates the cross talk between breast and glial cells in brain metastases leading to rapid disease progression.

doi: 10.18632/oncotarget.14047

Figure Lengend Snippet: Figure 7: Effects of glial cell cocultures on cytokine secretion. Control and PTEN overexpressing MDA-MB-231 BR cells were seeded in transwell inserts and cocultured for 48 h with astrocytes (A) or microglia (B). Antibody-spotted membranes were incubated with conditioned media of these cultures and levels of secreted proteins were visualized by HRP-conjugated antibodies. Proteins with a more than 2-fold expression difference are shown by open boxes. Results were verified by qPCR analyses (C). Astrocyte coculture reduced EGF and GM-CSF expression significantly in PTEN overexpressing (PTEN CoA) compared to control cells (CTL CoA, C). Astrocytes express the GM- CSF receptor (CSFR2A) only when cocultured with tumor cells. Microglia coculture reduced VEGFA expression in PTEN overexpressing (PTEN CoMG) compared to control cells (CTL CoMG). Student’s t-test was used to analyze statistical significance. *p < 0.05.

Article Snippet: Cytokine secretion was estimated from CM by Proteome ProfilerTM Antibody Array (Human XL Cytokine Array Kit, ARY022, R&D Systems, Wiesbaden, Germany) according to manufacturer’s instruction.

Techniques: Control, Incubation, Expressing

Journal: British Journal of Cancer

Article Title: Antibody targeting of CD24 efficiently retards growth and influences cytokine milieu in experimental carcinomas

doi: 10.1038/bjc.2013.102

Figure Lengend Snippet: Effects of SWA11 mAb treatment on intratumoural cytokine milieu in A549 tumours. ( A ) The levels of 40 different cytokines were determined in tumour lysates using a cytokine protein arrays. The level of cytokines was quantified using ImageJ software. Representative arrays from two independent SWA11 mAb treatment experiments are shown. ( B ) Protein levels of CCL5/RANTES, CXCL9/MIG and CCL2/MCP-1 in tumour lysates ( n =5 per group) were determined by ELISA. Data are normalised to the total protein concentration in tumour lysates. * P <0.05.

Article Snippet: Relative levels of 40 different cytokines and chemokines in xenograft tumour lysates were evaluated using a mouse cytokine antibody array kit (Proteome Profiler, R&D Systems) according to the manufacturer's recommendations.

Techniques: Software, Enzyme-linked Immunosorbent Assay, Protein Concentration

Journal: British Journal of Cancer

Article Title: Antibody targeting of CD24 efficiently retards growth and influences cytokine milieu in experimental carcinomas

doi: 10.1038/bjc.2013.102

Figure Lengend Snippet: Effects of SWA11 mAb treatment on intratumoural cytokine milieu in SKOV3ip tumours. ( A ) The levels of 40 different cytokines were determined in tumour lysates using cytokine protein arrays. The level of cytokines was quantified using ImageJ software. Representative arrays from two independent SWA11 mAb treatment experiments are shown. ( B ) Protein levels of CCL5/RANTES, CXCL9/MIG and CCL2/MCP-1 in tumour lysates ( n =5–6 per group) were determined by ELISA. Data are normalised to the total protein concentration in tumour lysates.

Article Snippet: Relative levels of 40 different cytokines and chemokines in xenograft tumour lysates were evaluated using a mouse cytokine antibody array kit (Proteome Profiler, R&D Systems) according to the manufacturer's recommendations.

Techniques: Software, Enzyme-linked Immunosorbent Assay, Protein Concentration

Journal: British Journal of Cancer

Article Title: Antibody targeting of CD24 efficiently retards growth and influences cytokine milieu in experimental carcinomas

doi: 10.1038/bjc.2013.102

Figure Lengend Snippet: Effects of combined SWA11 mAb and gemcitabine treatment on A549 tumour growth and intratumoural cytokine milieu. ( A ) SCID beige mice with established xenograft A549 lung carcinomas received treatment with SWA11 mAb (10 mg kg −1 ) or IgG 2A (10 mg kg −1 ) followed by gemcitabine at a dose of 12 mg kg −1 (10% of maximal tolerated dose) 1 day later ( n =5 per group). Control animals received SWA11 mAb (10 mg kg −1 ), IgG 2A (10 mg kg −1 ) or gemcitabine (12 mg kg −1 ) alone ( n =5 per group). Treatment was repeated three times with a 5-day interval. External size of A549 tumours was measured with a caliper. * P <0.05. Note that part of the results from this experiment are also shown in for illustration. ( B ) Effects of SWA11 mAb and gemcitabine treatment on the levels of 40 different cytokines were determined in A549 tumour lysates using a cytokine protein arrays. The level of cytokines was quantified using ImageJ software.

Article Snippet: Relative levels of 40 different cytokines and chemokines in xenograft tumour lysates were evaluated using a mouse cytokine antibody array kit (Proteome Profiler, R&D Systems) according to the manufacturer's recommendations.

Techniques: Control, Software

Journal: Physiological Reports

Article Title: Tuberous sclerosis complex exhibits a new renal cystogenic mechanism

doi: 10.14814/phy2.13983

Figure Lengend Snippet: List of antibodies used in the study

Article Snippet: Rabbit polyclonal Anti‐NCC , 1:60 , Stressmarq Biosciences; Victoria, BC, Canada , SPC‐402.

Techniques: Plasmid Preparation, Generated

Journal: PLoS ONE

Article Title: Targeted Deletion of HIF-1α Gene in T Cells Prevents their Inhibition in Hypoxic Inflamed Tissues and Improves Septic Mice Survival

doi: 10.1371/journal.pone.0000853

Figure Lengend Snippet: A: T lymphocytes deficient in HIF-1α undergo more cell divisions as compared to wild type T cells. Splenic T cells were purified, stained using CFSE, activated for 72 hours and analyzed by flow cytometry as described. A representative dot plot of wild type CD4 T cells (A) and HIF-1α deficient T cells (B) showing both activation by CD25 expression and cell divisions. (C) Comparison of number of divisions by CD4 T cells.*: p<0.05 vs. WT, N = 3 per group. B: T cell specific disruption of HIF-1 α gene substantially increases pro-inflammatory cytokine secretion by ex vivo TCR-activated T cells. Spleen T cells were derived from T cell lineage specific HIF-1 α deficient mice. Cells were activated for 24 h under hypoxic conditions (1% O 2 ). Extracellularly secreted cytokines were determined by ELISA. *:p<0.05 vs. WT, N = 5. C: Higher intracellular levels of IFN-γ production by inflamed peritoneum-located hypoxic HIF-1 α deficient CD8+ T cells as compared with similarly located in vivo hypoxic HIF-1 α expressing T cells after CLP . Peritoneal lavage was performed 72 hrs after CLP and 1,5×10 6 T cells were restimulated and stained with anti-IFN- γ mAb. D: Levels of proinflammatory cytokines TNF-α and IL-6 are significantly higher as compared to mice with selective disruption of HIF-1α gene in T-cells. Peritoneal fluid (TNF-α, left panel) and serum (IL-6, right panel) were withdrawn at the indicated times after CLP and cytokines were determined by ELISA. Closed circles: HIF-1α KO, open circles: WT. *:p<0.05 vs. WT, N = 4

Article Snippet: Intracellular staining of TCR-activated cells to evaluate IFN-γ production in situ was performed in permeabilized cells (after 4 h of anti-CD3 and anti-CD28 mAb restimulation) by flow cytometry analysis using anti-cytokine mAB (BD Pharmingen), 2 µM (final concentration) monensin (Cal-Biochem) and saponin buffer (PBS containing 0,1% (w/v) saponin, 0,1% BSA, 0,01 M HEPES and 0,1% sodium azide) as described Extracellular cytokine concentrations from serum, peritoneal fluid and from cell culture supernatant were determined by ELISA kits (BD Pharmingen) according to the manufacturer's instructions.

Techniques: Purification, Staining, Flow Cytometry, Activation Assay, Expressing, Ex Vivo, Derivative Assay, Enzyme-linked Immunosorbent Assay, In Vivo