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: Chinese Medicine

Article Title: Matrine improves bile acid metabolism and reduces inflammatory and oxidative stress in colitis via the JAK2 pathway

doi: 10.1186/s13020-026-01387-z

Figure Lengend Snippet: Effects of matrine on intestinal pathology, inflammatory function, and JAK2 expression in experimental colitis mice Dextran Sulfate Sodium (DSS)-induced murine model of colitis was established in mice; matrine and 5-ASA treatment was administered as described. A – C Body weight and colon length were measured. D , E Histopathological alterations in colon tissues were evaluated using H & E staining; DAI scores were assigned according to the staining. F , G The levels of IL-1β, TNF-α, IL-6, MPO, NO, and MDA in colon tissues were examined using commercial assay kits. H , I The protein levels of JAK2 in mice colon tissues were determined using Immunohistochemical staining (IHC staining) and Immunoblotting. Magnification = 100 × or 200 × for IHC staining. J The protein levels of p-JAK2, p-STAT3, and STAT3 were detected using Immunoblotting. **p < 0.01, vs. PBS group; #p < 0.05, ##p < 0.01 vs. DSS group

Article Snippet: Membrane was blocked within Odyssey blocking buffer (LI-COR Bioscience, Lincoln, USA) for 1 h at room temperature (RT), and then incubated with primary antibodies against p-JAK2 (AP0917, Abclonal, Woburn, USA), JAK2 (AF6022, Affinitiy Bioscience, Changzhou, China), MRP3 (bs-0656R, Bioss, Beijing, China), MRP4 (DF6921, Affinity Bioscience), IL-1β [12242, Cell Signaling Technology (CST), Danvers, USA], TNF-α (11948, CST), IL-6 (CSB-PA06757A0RB, CUSABIO, Wuhan, China), p-STAT3 (AF3293, Affinity Bioscience), STAT3 (10253-2-AP, Proteintech, Wuhan, China), p-STAT1 (28977-1-AP, Protientech), STAT1 (10144-2-AP, Proteintech), FXR (M022312, Abmart, Shanghai, China), and GAPDH (endogenous control, 60004-1-Ig, Proteintech) overnight at 4 °C (dilution 1:1000).

Techniques: Expressing, Staining, Immunohistochemical staining, Immunohistochemistry, Western Blot

Journal: Chinese Medicine

Article Title: Matrine improves bile acid metabolism and reduces inflammatory and oxidative stress in colitis via the JAK2 pathway

doi: 10.1186/s13020-026-01387-z

Figure Lengend Snippet: Effects of JAK2 overexpression on intestinal epithelial cells upon inflammation MODE-K cells were transfected with a JAK2-overexpressing plasmid (JAK2 oe) or a control vector, with or without subsequent LPS stimulation. A The protein levels of JAK2 and p-JAK2 and the inflammatory factors IL-1β, TNF-α, and IL-6 were determined by Immunoblotting. B Cellular levels of MPO, NO, and MDA were measured using biochemical kits. C Intracellular ROS production was assessed by DCFH-DA staining and quantified with flow cytometry. D The protein levels of the bile acid metabolism-related factors FXR, MRP3 and MRP4 were examined by Immunoblotting. E The mRNA levels of MRP3, MRP4, OSTα, and OSTβ were measured by qRT-PCR. **p < 0.01 vs. Vector group; ##p < 0.01 vs. Vector group; &&p < 0.01 vs. LPS + Vector group

Article Snippet: Membrane was blocked within Odyssey blocking buffer (LI-COR Bioscience, Lincoln, USA) for 1 h at room temperature (RT), and then incubated with primary antibodies against p-JAK2 (AP0917, Abclonal, Woburn, USA), JAK2 (AF6022, Affinitiy Bioscience, Changzhou, China), MRP3 (bs-0656R, Bioss, Beijing, China), MRP4 (DF6921, Affinity Bioscience), IL-1β [12242, Cell Signaling Technology (CST), Danvers, USA], TNF-α (11948, CST), IL-6 (CSB-PA06757A0RB, CUSABIO, Wuhan, China), p-STAT3 (AF3293, Affinity Bioscience), STAT3 (10253-2-AP, Proteintech, Wuhan, China), p-STAT1 (28977-1-AP, Protientech), STAT1 (10144-2-AP, Proteintech), FXR (M022312, Abmart, Shanghai, China), and GAPDH (endogenous control, 60004-1-Ig, Proteintech) overnight at 4 °C (dilution 1:1000).

Techniques: Over Expression, Transfection, Plasmid Preparation, Control, Western Blot, Staining, Flow Cytometry, Quantitative RT-PCR

Journal: Chinese Medicine

Article Title: Matrine improves bile acid metabolism and reduces inflammatory and oxidative stress in colitis via the JAK2 pathway

doi: 10.1186/s13020-026-01387-z

Figure Lengend Snippet: Effects of matrine on intestinal epithelial cell function upon inflammation Mouse intestinal epithelial cell line, MODE-K, was stimulated with LPS with or without matrine treatment (1, 2, 3 mg/ml), and examined for the protein levels of IL-1β, TNF-α, and IL-6 using Immunoblotting ( A ); The levels of MPO, NO, and MDA using commercial kits ( B ); Intracellular ROS levels were detected by flow cytometry ( C ); The protein levels of FXR, MRP3 and MRP4 were examined using Immunoblotting ( D ); The mRNA levels of MRP3, MRP4, OSTα and OSTβ were examined using qRT-PCR ( E ); The protein levels of p-JAK2, JAK2, p-STAT3, STAT3, p-STAT1, and STAT1 using Immunoblotting ( F ). **p < 0.01, vs. PBS group; #p < 0.05, ##p < 0.01 vs. LPS group

Article Snippet: Membrane was blocked within Odyssey blocking buffer (LI-COR Bioscience, Lincoln, USA) for 1 h at room temperature (RT), and then incubated with primary antibodies against p-JAK2 (AP0917, Abclonal, Woburn, USA), JAK2 (AF6022, Affinitiy Bioscience, Changzhou, China), MRP3 (bs-0656R, Bioss, Beijing, China), MRP4 (DF6921, Affinity Bioscience), IL-1β [12242, Cell Signaling Technology (CST), Danvers, USA], TNF-α (11948, CST), IL-6 (CSB-PA06757A0RB, CUSABIO, Wuhan, China), p-STAT3 (AF3293, Affinity Bioscience), STAT3 (10253-2-AP, Proteintech, Wuhan, China), p-STAT1 (28977-1-AP, Protientech), STAT1 (10144-2-AP, Proteintech), FXR (M022312, Abmart, Shanghai, China), and GAPDH (endogenous control, 60004-1-Ig, Proteintech) overnight at 4 °C (dilution 1:1000).

Techniques: Cell Function Assay, Western Blot, Flow Cytometry, Quantitative RT-PCR

Journal: Chinese Medicine

Article Title: Matrine improves bile acid metabolism and reduces inflammatory and oxidative stress in colitis via the JAK2 pathway

doi: 10.1186/s13020-026-01387-z

Figure Lengend Snippet: Dynamic effects of matrine and JAK2 on intestinal epithelial cell function upon inflammation MODE-K cells were stimulated with LPS with or without matrine treatment (2 mg/ml), transfected with JAK2-overexpressing plasmid (JAK2 oe), and examined for the protein levels of IL-1β, TNF-α, and IL-6 using Immunoblotting ( A ); the levels of MPO, NO, and MDA using commercial kits ( B ); Intracellular ROS levels were detected by flow cytometry ( C ); The protein levels of FXR, MRP3 and MRP4 were examined using Immunoblotting ( D ); The mRNA levels of MRP3, MRP4, OSTα and OSTβ were examined using qRT-PCR ( E ); the protein levels of JAK2, p-STAT3, STAT3, p-STAT1, and STAT1 using Immunoblotting ( F ). **p < 0.01, vs. LPS group; #p < 0.05, ## p < 0.01 vs. LPS + matrine + vector group

Article Snippet: Membrane was blocked within Odyssey blocking buffer (LI-COR Bioscience, Lincoln, USA) for 1 h at room temperature (RT), and then incubated with primary antibodies against p-JAK2 (AP0917, Abclonal, Woburn, USA), JAK2 (AF6022, Affinitiy Bioscience, Changzhou, China), MRP3 (bs-0656R, Bioss, Beijing, China), MRP4 (DF6921, Affinity Bioscience), IL-1β [12242, Cell Signaling Technology (CST), Danvers, USA], TNF-α (11948, CST), IL-6 (CSB-PA06757A0RB, CUSABIO, Wuhan, China), p-STAT3 (AF3293, Affinity Bioscience), STAT3 (10253-2-AP, Proteintech, Wuhan, China), p-STAT1 (28977-1-AP, Protientech), STAT1 (10144-2-AP, Proteintech), FXR (M022312, Abmart, Shanghai, China), and GAPDH (endogenous control, 60004-1-Ig, Proteintech) overnight at 4 °C (dilution 1:1000).

Techniques: Cell Function Assay, Transfection, Plasmid Preparation, Western Blot, Flow Cytometry, Quantitative RT-PCR

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Super enhancer-driven LINC01013 mediates hypoxia-induced mitochondrial dysfunction by HSPA9 to determine pulmonary arterial smooth muscle cell fate

doi: 10.1007/s00018-025-06071-3

Figure Lengend Snippet: SE-associated LINC01013 was transcriptionally activated by CEBPB in hPASMCs. A Prediction of candidate transcription factors and binding sites. (Transcription factor related databases: JASPAR: https://jaspar.elixir.no/ ; PROMO: https://alggen.lsi.upc.es/cgi-bin/promo_v3/promo/promoinit.cgi?dirDB=TF_8.3 ; GENECARD: https://www.genecards.org/ ; AnimalTFDB: http://bioinfo.life.hust.edu.cn/HumanTFDB/#!/ ; Super enhancer related database: LncSEA: https://bio.liclab.net/LncSEA/ ). B The schematic diagram illustrates the LINC01013 promoter, divided into segments P1-P4, and the binding sites of transcription factors. C Four constituents (E1-E4) of SE region of LINC01013 derived from the WashU Epigenome Browser databases ( http://epigenomegateway.wustl.edu/browser/ ). D , E hPASMCs were subjected to ChIP analysis using antibodies against H3K27ac, H3K4me1 and CEBPB. The association with the SE region (D, E1-E4) and promoter region (E, P1-P4) of LINC01013 was quantified by RT‒qPCR ( n = 3). F hPASMCs were treated with CEBPB siRNA and subjected to ChIP analysis using antibodies against H3K27ac. The association with the E2 of SE (left) and P1-P3 promoter regions (right) of LINC01013 was quantified by RT-qPCR ( n = 3). G Schematic diagram of transcribing LINC01013 in hPASMCs. All values are presented as the mean ± SEM. Statistical analysis was performed with one-way ANOVA or Student’s t-test. * p < 0.05, ** p < 0.01, *** p < 0.001. Nor, normoxia; Hyp, hypoxia; NC, negative control; IP, immunoprecipitation; IgG, Immunoglobulin G; TSS, transcription initiation site

Article Snippet: The antibody against CEBPB (PB9171, BA0670, 1:500, Boster, Wuhan, China), H3K27ac (A7253, 1:500, ABclonal, Wuhan, China), H3K4me1 (A2355, 1:500, Wuhan, China), PCNA (A00125, 1:500, Boster, Wuhan, China), Cyclin A (PB0515, 1:500, Boster, Wuhan, China), Cyclin D (BM4272, 1:500, Boster, Wuhan, China), IL-6 (AF7236, 1:500, Beyotime, Shanghai, China), TNF-α (AF8208, 1:500, Beyotime, Shanghai, China), PKM2 (4053, 1:1000, Cell Signaling, MA, US), HK II (66974-1-Ig, 1:1000, Proteintech, IL, USA), PDH (2784, 1:1000, Cell Signaling, MA, US), HSPA9 (14887-1-AP, 1:5000, Proteintech, IL, USA), VDAC1 (10866-1-AP, 1:5000, Proteintech, IL, USA), and β-actin (TA-09, 1:1000, ZSGB‐BIO, Beijing, China) was incubated at 4 °C overnight, followed by incubation with appropriate horseradish peroxidase-conjugated secondary antibodies at room temperature for 1 h, and proteins were visualized with enhanced chemiluminescence reagents.

Techniques: Binding Assay, Derivative Assay, Quantitative RT-PCR, Negative Control, Immunoprecipitation