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: