Journal: Leukemia

Article Title: HLXB9 activates IL6 in Hodgkin lymphoma cell lines and is regulated by PI3K signalling involving E2F3.

doi: 10.1038/sj.leu.2403716

Figure Lengend Snippet: Figure 3 Expression of IL6 and HLXB9 by RT-PCR in response to HLXB9 modulation. (a) Western blot control of expression vector pcrHLXB9 in comparison to empty vector pcDNA3 transfected in 293 cells. GDM-1 positive control cells strongly express HLXB9.29 (b) HL cell line L-540 was treated with 107 M TPA or 0.1% DMSO in combination with transfection of pcrHLXB9. Expression of IL6 is detectable after TPA treatment only. Transfection with pcrHLXB9 enhances expression of IL6. (c) HDLM-2 cells were treated with antisense-oligo (AS) directed against HLXB9 in comparison to control- oligo (AS-control). Both HLXB9 and IL6 evidenced reduced expres- sion. NTC: no template control.

Article Snippet: Inhibition of secreted IL6 protein was performed using anti-IL6 antibody (R&D Systems, Wiesbaden, Germany) and for stimulation experiments IL6 protein was purchased from Strathmann Biotec AG (Hamburg, Germany).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Control, Plasmid Preparation, Comparison, Transfection, Positive Control

Journal: Leukemia

Article Title: HLXB9 activates IL6 in Hodgkin lymphoma cell lines and is regulated by PI3K signalling involving E2F3.

doi: 10.1038/sj.leu.2403716

Figure Lengend Snippet: Figure 4 Schematic diagram of IL6 promoter constructs. IL6 promoter sequences were cloned before a reporter gene (EGFP). Potential binding sites for AP-1, NFkB, and GBX2/HLXB9 (EHG) are indicated.

Article Snippet: Inhibition of secreted IL6 protein was performed using anti-IL6 antibody (R&D Systems, Wiesbaden, Germany) and for stimulation experiments IL6 protein was purchased from Strathmann Biotec AG (Hamburg, Germany).

Techniques: Construct, Clone Assay, Binding Assay

Journal: Cancer Research

Article Title: Slit2 Inhibits Breast Cancer Metastasis by Activating M1-Like Phagocytic and Antifibrotic Macrophages

doi: 10.1158/0008-5472.can-20-3909

Figure Lengend Snippet: Figure 5. Slit2 reduces the expression of IL6, thereby inhibiting TAMs activity. A, Total RNA of CD11bþ cells sorted from rSlit2-N PBS-treated MMTV-PyMT tumorswas subjected to gene expression analysis using NanoString technology. The heatmap shows differentially expressed top genes. B, Total RNA was isolated from MDA-MB-231 cells overexpressing Slit2 (231-Slit2) or vector control (231-vec) and analyzed for gene expression using microarray technology. The heatmap shows differentially expressed top genes. C, 231-Slit2 or 231-Vec cells CM was subjected to cytokine array analysis. The representative image shows differentially expressed molecules. D, The graph shows levels of IL6 in CM derived from 231-Sli2 or Vec CM detected by ELISA. E, Expression of Robo1 in MDA-MB-231 cells transduced with lentivirus expressing siRNA specific to Robo1 (si-Robo1) or control (si-ctrl) by Western blot. F, b-actin was used as loading control. MDA-MB-231 parental control, si-Robo1, or si-ctrl from E was treated with rSlit2-N or PBS and phosphorylation at p65 of NFkB (p-NFkB) or total NFkB (t-NFkB) was analyzed. G, Graph showing levels of IL6 in si-Robo1 or si-ctrl MDA-MB-231 cells treated with rSlit2-N or PBS detected by ELISA (n ¼ 3 each group). H, BMDMs were pretreated with serum-free CM (CTRL), or 231-Slit2 CM or 231-Vec CM or 231-Vec CM preincubated with IL6 nAb (231-Vec þ IL6nAb) or IL6 nAb. After 2 hours, pH-Rhodo–labeled S. Aureus particles were added to the cells and recombinant IL6 was added to the 231-Slit2 CM cells (231-Slit2-rIL6). The kinetics of S. Aureus phagocytosis was analyzed using a fluorescence plate reader at every 15 minutes up to 4 hours. , P < 0.05; , P < 0.01; , P < 0.001 using Student t test.

Article Snippet: The cells were pretreated with mouse rSlit2-N (100 ng/mL) or IL6 neutralization antibody (BioXCell# BE0046) for 2 hours.

Techniques: Expressing, Activity Assay, Gene Expression, Isolation, Plasmid Preparation, Control, Microarray, Derivative Assay, Enzyme-linked Immunosorbent Assay, Transduction, Western Blot, Phospho-proteomics, Labeling, Recombinant

Journal: medRxiv

Article Title: Biofabrication of multiplexed electrochemical immunosensors for simultaneous detection of clinical biomarkers in complex fluids

doi: 10.1101/2022.03.18.22272576

Figure Lengend Snippet: Calibration curves for different biomarkers using antifouling nanocomposite coated EC Biosensors. The left y-axis shows the current intensity for different concentrations of biomarkers run on EC biosensors using unprocessed human plasma. Different biomarkers tested include a) IL-6, b) IL-8, c) IL-8, d) VEGF, e) Ag85B, and f) PCT. Error bars represent the s.d. of the mean; n = 3. Analysis was done using 4-Parameter Logistic (4PL) curve fitting.

Article Snippet: Detection antibodies used for the assay include anti-BNP (HyTest, no. 24C5cc), anti-NT-proBNP (Medix Biochemica, no. 100 712), anti-cTnI (Abcam, no. ab243982), anticTnI-TC (Advanced ImmunoChemical, USA no. 2-TC), anti-GFAP (HyTest, no. GFAP81cc), anti-S-100b (HyTest, no. 6G1cc), anti NF-L (Uman Diagnostics, no. 27018), anti-IL-6, anti-IL-8, anti-IL-18, anti-VEGF, and anti-Ag85-B, respectively.

Techniques:

Journal: EBioMedicine

Article Title: Ketogenesis acts as an endogenous protective programme to restrain inflammatory macrophage activation during acute pancreatitis

doi: 10.1016/j.ebiom.2022.103959

Figure Lengend Snippet: βOHB limits proinflammatory macrophage activation via class I HDACs, (a, b) Gene set enrichment analysis (GSEA) of gene expression in murine BMDMs are depicted. (a) Enrichment of TSA-upregulated signature genes (GSE22049) in βOHB-treated BMDMs. (b) Enrichment of HDAC3 depletion related signature genes (GES33162) in βOHB-treated BMDMs. (c, d) BMDMs treated with LPS (100 ng/ml) together with βOHB (10 mM), TSA (1 μΜ), or MS-275 (20 μM) for 4 h. (c) Flow cytometry analysis of intracellular TNFα levels. (d) Quantitative PCR analysis of Nos2, Il6, Il12b , and Tnfa mRNA expression. (e) ChIP-qPCR analysis of H3Ac and H4Ac occupancy in Edn1, Il12a , and Nos2 loci in the indicated group, with IgG as the control. (f) Ingenuity Pathway Analysis (IPA) of genes in cluster A. (g) BMDMs were pretreated with βOHB (10 mM) or TSA (1 μΜ) 1 h prior to LPS (100 ng/ml) stimulation for 15 min. Acylated p65 levels were determined by immunoprecipitation, while p-p65 and total p65 were determined by immunoblotting. (h) Mice were i.p. injected with TSA (0.1 mg/kg), MS-275 (20 mg/kg) or DMSO as a control 1 h prior to the CER-AP (Cer*7) induction and were harvested at 24 h thereafter (n = 5 / group). Frequency of TNFα + macrophagess (F4/80 + CD11b + ) in pancreatic leukocytes from the indicated groups. Data are presented as the mean ± SD in bar graphs (c–e) or the mean ± SEM in dot plots (h). *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: For treatments, the Balb/c mice were (1) fed with 1,3-butanediol (5.72% (w/v); Sigma-Aldrich) or normal water (control) one week before disease induction; received a single intraperitoneal injection of βOHB (3 mmol/kg; Sigma-Aldrich) or normal saline at the indicated time; and (3) the CPT1α inhibitor etomoxir (20 mg/kg, Sigma-Aldrich), histone deacetylase (HDAC) pan inhibitor TSA (0.1 mg/kg; Targetmol), HDAC1/3 inhibitor MS-275 (20 mg/kg; Targetmol), or dimethyl sulfoxide (DMSO) as a control 1 h before disease induction.

Techniques: Activation Assay, Gene Expression, Flow Cytometry, Real-time Polymerase Chain Reaction, Expressing, ChIP-qPCR, Control, Immunoprecipitation, Western Blot, Injection