Journal: BMC Cancer

Article Title: IL6 secreted by Ewing sarcoma tumor microenvironment confers anti-apoptotic and cell-disseminating paracrine responses in Ewing sarcoma cells

doi: 10.1186/s12885-015-1564-7

Figure Lengend Snippet: Serum IL6 levels at initial diagnosis of ES in 12 patients measured by ELISA

Article Snippet: Recombinant human IL6 (rhIL6, Cat. No 206-IL) and human anti-IL6R antibody (clone 17506, Cat. No MAB227) were purchased from R&D Systems, Minneapolis, USA.

Techniques: Biomarker Discovery

Journal: BMC Cancer

Article Title: IL6 secreted by Ewing sarcoma tumor microenvironment confers anti-apoptotic and cell-disseminating paracrine responses in Ewing sarcoma cells

doi: 10.1186/s12885-015-1564-7

Figure Lengend Snippet: a IL6R and IL6ST mRNA expression of seven ES tumor samples by RT-PCR. IFN-γ treated PBMC were used as positive control (left lane). All tumor specimens expressed both IL6 receptor subunits. b IL6R and IL6ST mRNA expression in nine ES cell lines (RT-PCR). IFN-γ treated PBMC were used as positive controls (left lane). c Assessment of IL6 receptor complex cell surface expression by flow cytometry. The prostate cancer cell line LNCaP was used as a positive control. IL6R and IL6ST cell surface staining of 4 additional ES cell lines is shown in Additional file : Figure S1

Article Snippet: Recombinant human IL6 (rhIL6, Cat. No 206-IL) and human anti-IL6R antibody (clone 17506, Cat. No MAB227) were purchased from R&D Systems, Minneapolis, USA.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Positive Control, Flow Cytometry, Staining

Journal: BMC Cancer

Article Title: IL6 secreted by Ewing sarcoma tumor microenvironment confers anti-apoptotic and cell-disseminating paracrine responses in Ewing sarcoma cells

doi: 10.1186/s12885-015-1564-7

Figure Lengend Snippet: a p-STAT3 phospho-specific immunoblot of four ES cell lines. rhIL6-mediated STAT3 activation was detected after 20 min incubation of indicated IL6 concentrations in cell lines A4573, TC32 and CHP. All cell lines showed low intrinsic phosphorylation of STAT3. Cell line 5838 did not display an increase in p-STAT3 after incubation with rhIL6. b p-STAT3 phospho-specific immunoblot of cell line A4573 after incubation with 50 ng/ml rhIL6 for indicated time periods. Maximal levels of phosphorylated STAT3 were seen 20 min after adding rhIL6. STAT3 phosphorylation decreased after 2 h of incubation. c Phosphorylation of STAT3 in A4573 ES cell line is IL6-specific. Cells were stimulated with rhIL6 (100 ng/ml for 20 min). An IL6 receptor-specific antagonistic antibody (2 μg/ml) was added 30 min prior to rhIL6 incubation where indicated. The prostate cancer cell line LNCaP was used as a positive control. STAT3 phosphorylation mediated by rhIL6 could be blocked by an anti-IL6 receptor antibody. Data representative for three independent protein harvests and immunoblots

Article Snippet: Recombinant human IL6 (rhIL6, Cat. No 206-IL) and human anti-IL6R antibody (clone 17506, Cat. No MAB227) were purchased from R&D Systems, Minneapolis, USA.

Techniques: Western Blot, Activation Assay, Incubation, Phospho-proteomics, Positive Control

Journal: BMC Cancer

Article Title: IL6 secreted by Ewing sarcoma tumor microenvironment confers anti-apoptotic and cell-disseminating paracrine responses in Ewing sarcoma cells

doi: 10.1186/s12885-015-1564-7

Figure Lengend Snippet: a Increased viability of serum-starved A4573 ES cells by repetitive stimulation with rhIL6 (d 0, 2, 4; 10 ng/ml). Fold-change (x) of MTT conversion rates (n = 5; mean ± SD) relative to rates of cells measured 24 h after seeding and set to 1 (100 %) are shown. Statistical significance of IL6 treatment relative to controls is indicated (Mann–Whitney- U -Test; * = p < 0.05). b Serial supply of rhIL6 reduces rate of apoptosis from serum starvation in A4573 ES cells. Serum-starved cells of ES cell line A4573 cells were stimulated repetitively with rhIL6 (10 ng/ml on indicated days) and harvested on day 5. Sub-G1 DNA content was used as a measure of apoptosis. Bars (n = 3; mean ± SD) indicate fraction of cells (%) containing a sub-G1 DNA content. Similar results could be shown in two other independent experiments

Article Snippet: Recombinant human IL6 (rhIL6, Cat. No 206-IL) and human anti-IL6R antibody (clone 17506, Cat. No MAB227) were purchased from R&D Systems, Minneapolis, USA.

Techniques: MANN-WHITNEY

Journal: BMC Cancer

Article Title: IL6 secreted by Ewing sarcoma tumor microenvironment confers anti-apoptotic and cell-disseminating paracrine responses in Ewing sarcoma cells

doi: 10.1186/s12885-015-1564-7

Figure Lengend Snippet: a Immunofluorescence of A4573/GFP ES cells grown on a CAM and treated with single IL6 dose as indicated. b Cell coverage across equal-size zones 3, 2 and 1 (right) was estimated by superimposed 3 × 12 square area with values ranging from 0 (no cells present) to 5 (square 100 % filled with cells). See for details. The bar graph (left) illustrates relative mean (± SEM) coverage values for zones 3, 2 and 1 from a total of three independent experiments as a measure of cell motility in response to same-volume applications of PBS, 250 ng and 500 ng rhIL6. Addition of rhIL6 dose-dependently and significantly (* = p < 0.05) increased motility of A4573 ES cells on the surface of the CAM from zone 3 (explant perimeter) to zone 1 (furthest distance)

Article Snippet: Recombinant human IL6 (rhIL6, Cat. No 206-IL) and human anti-IL6R antibody (clone 17506, Cat. No MAB227) were purchased from R&D Systems, Minneapolis, USA.

Techniques: Immunofluorescence

Journal: BMC Cancer

Article Title: IL6 secreted by Ewing sarcoma tumor microenvironment confers anti-apoptotic and cell-disseminating paracrine responses in Ewing sarcoma cells

doi: 10.1186/s12885-015-1564-7

Figure Lengend Snippet: a Detection of IL6 mRNA expression in 7 ES tumor samples using RT-PCR. IFN-γ treated PBMC were used as positive control. 3 out of 7 tumor samples expressed IL6 mRNA. b IL6 mRNA expression in 9 ES cell lines by RT-PCR. IFN-γ treated PBMC were used as positive control. 4 out of 9 cell lines expressed IL6 mRNA

Article Snippet: Recombinant human IL6 (rhIL6, Cat. No 206-IL) and human anti-IL6R antibody (clone 17506, Cat. No MAB227) were purchased from R&D Systems, Minneapolis, USA.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Positive Control

Journal: BMC Cancer

Article Title: IL6 secreted by Ewing sarcoma tumor microenvironment confers anti-apoptotic and cell-disseminating paracrine responses in Ewing sarcoma cells

doi: 10.1186/s12885-015-1564-7

Figure Lengend Snippet: Concentration of IL6 in the supernatant of 5 ES cell lines cultured in vitro (ELISA)

Article Snippet: Recombinant human IL6 (rhIL6, Cat. No 206-IL) and human anti-IL6R antibody (clone 17506, Cat. No MAB227) were purchased from R&D Systems, Minneapolis, USA.

Techniques: Concentration Assay, Cell Culture, In Vitro, Enzyme-linked Immunosorbent Assay

Journal: BMC Cancer

Article Title: IL6 secreted by Ewing sarcoma tumor microenvironment confers anti-apoptotic and cell-disseminating paracrine responses in Ewing sarcoma cells

doi: 10.1186/s12885-015-1564-7

Figure Lengend Snippet: Immunohistochemical analysis of two ES tumor specimens and control tissues for IL6, vimentin and smooth muscle actin (SMA). Pictures A and B show staining of control tissues with anti-IL6-ab (both 200×): a Appendix: positive staining of cells/extracellular space of lamina propria. b Tonsil: positive staining of cells/extracellular space in center of lymphoid follicles. Pictures C and D: examples of ES, negative and positive for IL6. c1 and c2 negative for IL6 in connective tissue septa within the tumor (Pt. 8). d1 and d2 ) positive for IL6 in connective tissue septa within the tumor (Pt. 7). In both samples no staining could be observed in tumor tissue. c1 and d1 : 100×, c2 and d2 : 200×. Pictures e1 - e3 : comparison of different stainings of ES, positive case (Pt. 7), all 200×): e1 IL6 staining, showing positivity in connective tissue septa between negative (blue) tumor cells. e2 Staining against vimentin demonstrates mesenchymal character of both tumor cells and cells in connective tissue septa. e3 Smooth muscle actin highlights the cells within the connective tissue septa, defining them as fibroblasts

Article Snippet: Recombinant human IL6 (rhIL6, Cat. No 206-IL) and human anti-IL6R antibody (clone 17506, Cat. No MAB227) were purchased from R&D Systems, Minneapolis, USA.

Techniques: Immunohistochemical staining, Control, Staining, Comparison

Journal: iScience

Article Title: The long non-coding RNA CDK6-AS1 overexpression impacts on acute myeloid leukemia differentiation and mitochondrial dynamics

doi: 10.1016/j.isci.2021.103350

Figure Lengend Snippet:

Article Snippet: interleukin6 (rhIL-6) , Milteny Biotec , Cat# 130-093-931.

Techniques: Recombinant, Staining, Reverse Transcription, SYBR Green Assay, Purification, Reporter Assay, In Situ, Membrane, Expressing, Microarray, Luciferase, Plasmid Preparation, Software