Journal: International Journal of Cardiology

Article Title: Enhanced IL-17 signalling following myocardial ischaemia/reperfusion injury

doi: 10.1016/j.ijcard.2011.08.849

Figure Lengend Snippet: Primers sequences used for qPCR analysis.

Article Snippet: In a different set of experiments rats were treated with 200 μg of anti-IL-17A polyclonal antibody intraperitoneally (R & D Systems, AF-421-NA) or PBS 2 h before undergoing in vivo ischemia/reperfusion injury.

Techniques:

Journal: International Journal of Cardiology

Article Title: Enhanced IL-17 signalling following myocardial ischaemia/reperfusion injury

doi: 10.1016/j.ijcard.2011.08.849

Figure Lengend Snippet: IL-17A, IL-17F, IL-17 receptor and IL-17 target genes are transcriptionally upregulated following in vivo I/R injury. RNA and protein was extracted from the left ventricles of rats undergoing sham operation or 25 min ischaemia and 2 h reperfusion (I/R). (A) IL-17RA, IL-17A and IL-17F expression was measured by qPCR. (B) IL-17A and IL-17RA protein levels were assessed by western blot, GAPDH was used as a loading control. (C) Expression of the IL-17 target genes IL-6, IL-1, iNOS and MMP-9 was measured by qPCR.

Article Snippet: In a different set of experiments rats were treated with 200 μg of anti-IL-17A polyclonal antibody intraperitoneally (R & D Systems, AF-421-NA) or PBS 2 h before undergoing in vivo ischemia/reperfusion injury.

Techniques: In Vivo, Expressing, Western Blot, Control

Journal: International Journal of Cardiology

Article Title: Enhanced IL-17 signalling following myocardial ischaemia/reperfusion injury

doi: 10.1016/j.ijcard.2011.08.849

Figure Lengend Snippet: IL-17 induces Cxcl1, IL-6 and Socs3 expression in cardiac myocytes. Neonatal rat ventricular myocytes were treated with 10 ng/ml IL-17 for the indicated times and the levels of (A) Cxcl1, (B) IL-6, (C) MCP-1 and (D) Socs3 were measured by qPCR. Experiments were repeated in duplicate, statistical analysis was carried out using a one-way ANOVA followed by Dunnett's post test, *p < 0.05, **p < 0.01, ***p < 0.001. (E) I/R injury and IL-17 have additive effects on Cxcl1 expression. NRVMs were subjected to 4 h ischaemia and 6 h reperfusion with or without 10 ng/ml IL-17 which was added at the time of reperfusion. The expression of Cxcl1 was analysed by qPCR. Statistical analysis was carried out using student's t -test. **p < 0.01, **p < 0.001.

Article Snippet: In a different set of experiments rats were treated with 200 μg of anti-IL-17A polyclonal antibody intraperitoneally (R & D Systems, AF-421-NA) or PBS 2 h before undergoing in vivo ischemia/reperfusion injury.

Techniques: Expressing

Journal: International Journal of Cardiology

Article Title: Enhanced IL-17 signalling following myocardial ischaemia/reperfusion injury

doi: 10.1016/j.ijcard.2011.08.849

Figure Lengend Snippet: IL-17 mediated upregulation of Cxcl1 and IL-6 in cardiac myocytes is MAPK dependent. (A) NRVMs were treated with 10 ng/ml IL-17 for the indicated times and cell lysates were analysed by Western blot using the indicated antibodies. (B) NRVMs were pre-treated for 30 min with the indicated inhibitors followed by IL-17 stimulation for 6 h. The expression of Cxcl1 and IL-6 was measured by qPCR. Statistical analysis was carried out using a student's t -test. *p < 0.05, **p < 0.01, **p < 0.001.

Article Snippet: In a different set of experiments rats were treated with 200 μg of anti-IL-17A polyclonal antibody intraperitoneally (R & D Systems, AF-421-NA) or PBS 2 h before undergoing in vivo ischemia/reperfusion injury.

Techniques: Western Blot, Expressing

Journal: International Journal of Cardiology

Article Title: Enhanced IL-17 signalling following myocardial ischaemia/reperfusion injury

doi: 10.1016/j.ijcard.2011.08.849

Figure Lengend Snippet: IL-17 receptor and IL-17F are transcriptionally upregulated in cardiac myocytes. (A) NRVM were subjected to 4 h in vitro ischaemia and up to 24 h reperfusion in normal media after which the levels of IL-17 receptor and IL-17F were assessed by qPCR. (B) H9c2 cells were subjected to in vitro I/R for the indicated times and the levels of IL-17F and Cxcl1 were measured by qPCR. (C) NRVMs were treated with 200 μM H 2 O 2 for 6 h and the expression of the indicated genes was measured by qPCR. (D) NRMVs were transduced with STAT3C adenovirus at MOI = 100. After 48 h, increased expression of STAT3 was confirmed by Western blot and IL-17F expression was measured by qPCR. *p < 0.05, **p < 0.01, ***p < 0.001, student's t -test, n = 3 per group, repeated in duplicate.

Article Snippet: In a different set of experiments rats were treated with 200 μg of anti-IL-17A polyclonal antibody intraperitoneally (R & D Systems, AF-421-NA) or PBS 2 h before undergoing in vivo ischemia/reperfusion injury.

Techniques: In Vitro, Expressing, Transduction, Western Blot

Journal: International Journal of Cardiology

Article Title: Enhanced IL-17 signalling following myocardial ischaemia/reperfusion injury

doi: 10.1016/j.ijcard.2011.08.849

Figure Lengend Snippet: Blocking IL-17 signalling with a specific anti-IL-17 neutralizing antibody reduced creatine phosphokinase and apoptotic cell death in the myocardium following in vivo I/R injury. Plasma concentration of CPK (A) and apoptosis assessed by the TUNEL assay (B) in sham-operated (control) or rats exposed to in vivo I/R injury (I/R) or I/R plus treatment with IL-17 neutralizing antibody (IL-17 Bo Ab). Immunofluorescence data (left panel) (Data are mean of ± 6 rats **P < 0.001). Anti-IL-17 neutralizing antibody also reduced the increase in cleaved/active caspase-3 expression following in vivo I/R injury (C). Western blot analysis was performed on tissue lysates prepared as in Fig. 3 A above and immunoblotted with a specific antibody against the cleaved and active form of caspase-3 and GAPDH.

Article Snippet: In a different set of experiments rats were treated with 200 μg of anti-IL-17A polyclonal antibody intraperitoneally (R & D Systems, AF-421-NA) or PBS 2 h before undergoing in vivo ischemia/reperfusion injury.

Techniques: Blocking Assay, In Vivo, Clinical Proteomics, Concentration Assay, TUNEL Assay, Control, Immunofluorescence, Expressing, Western Blot

Journal: Respiratory Research

Article Title: CD8 positive T cells express IL-17 in patients with chronic obstructive pulmonary disease

doi: 10.1186/1465-9921-12-43

Figure Lengend Snippet: IL-17A and IL-17F expression in COPD patients . (A) Immunohistochemistry, positive staining appears brown color. Magnification, 100 ×. Scale bar = 50 μm. (B) IL-17F expression in epithelium of airways of COPD patients. IL-17F positive area in epithelium was measured as outlined in text. (C) IL-17A and IL-17F expression in submucosa of airways of COPD patients. Absolute IL-17A + and IL-17F + positive cells in submucosa were counted. Results are expressed as median (range), n = 15 and 16 subjects for controls and COPD patients respectively. *** P < 0.0001.

Article Snippet: Slides were incubated overnight at 4°C using diluted goat anti-human IL-17A (AF317-NA, R&D Systems) or IL-17F (AF1335, R&D Systems) polyclonal antibodies or relevant isotype controls (AB-108-C, R&D Systems).

Techniques: Expressing, Immunohistochemistry, Staining

Journal: Respiratory Research

Article Title: CD8 positive T cells express IL-17 in patients with chronic obstructive pulmonary disease

doi: 10.1186/1465-9921-12-43

Figure Lengend Snippet: Percentage of IL-17A + and IL-17F + cells in airway submucosal cells of COPD patients . (A) Submucosal cells in airways of COPD patients. (B) Percentage of IL-17A + and IL-17F + cells in airway submucosal cells of COPD patients. Results are expressed as median (range), n = 15 and 16 subjects for controls and COPD patients respectively. ** P < 0.001, *** P < 0.0001.

Article Snippet: Slides were incubated overnight at 4°C using diluted goat anti-human IL-17A (AF317-NA, R&D Systems) or IL-17F (AF1335, R&D Systems) polyclonal antibodies or relevant isotype controls (AB-108-C, R&D Systems).

Techniques:

Journal: Respiratory Research

Article Title: CD8 positive T cells express IL-17 in patients with chronic obstructive pulmonary disease

doi: 10.1186/1465-9921-12-43

Figure Lengend Snippet: IL-17A and IL-17F mRNA expression in airways of COPD patients . (A) Quantitative RT-PCR was performed from frozen airways sections of COPD patients. One representative example from 7 subjects with similar results is shown. (B) Quantification of IL-17A and IL-17F mRNA expression in airways of control subjects and COPD patients. Results are expressed as means ± SEM. N = 7 for both control subjects and COPD patients.

Article Snippet: Slides were incubated overnight at 4°C using diluted goat anti-human IL-17A (AF317-NA, R&D Systems) or IL-17F (AF1335, R&D Systems) polyclonal antibodies or relevant isotype controls (AB-108-C, R&D Systems).

Techniques: Expressing, Quantitative RT-PCR, Control

Journal: Respiratory Research

Article Title: CD8 positive T cells express IL-17 in patients with chronic obstructive pulmonary disease

doi: 10.1186/1465-9921-12-43

Figure Lengend Snippet: Double immunofluorescence staining for detection of IL-17A and IL-17F expression in CD4 + and CD8 + T cells in airways of COPD patients . (A) Double immunofluorescence staining was performed. Scale bar = 5 μm. (B) Percentage of CD4 + and CD8 + T cells that express IL-17A and IL-17F. Results are expressed as means ± SEM. (C) Percentage of CD4 + and CD8 + T cells that express IL-17A and IL-17F in total IL-17A + and IL-17F + cells. Results are expressed as means ± SEM. *P < 0.05. N = 3 COPD patients. (D) IL-17A and IL-17F mRNA expression in CD8 + T cells in airways of COPD patients. Immunohistochemistry determined CD8 + T cells were selected by LCM, and then RT-PCR was performed to detect the mRNA expression of IL-17A and IL-17F. One representative result from 3 subjects is shown.

Article Snippet: Slides were incubated overnight at 4°C using diluted goat anti-human IL-17A (AF317-NA, R&D Systems) or IL-17F (AF1335, R&D Systems) polyclonal antibodies or relevant isotype controls (AB-108-C, R&D Systems).

Techniques: Double Immunofluorescence Staining, Expressing, Immunohistochemistry, Reverse Transcription Polymerase Chain Reaction

Journal: PLoS ONE

Article Title: Microbial Dysbiosis in Colorectal Cancer (CRC) Patients

doi: 10.1371/journal.pone.0016393

Figure Lengend Snippet: Samples from the same individuals and colonic sites were submitted to DNA extraction and PCR. Interleukin 17 (IL17)-immunoreactive cells in colonic tissues were mainly located in the lamina propria in the normal tissues [A: colonic normal mucosa from a normal individual (high magnification x40 at the bottom), B: colonic normal mucosa from a patient with colon cancer (high magnification x40 at the bottom)] and infiltrated the tumour tissue in a the same individual than in B [C: IL17 imlmunoreactive cells infiltrating the tumour with high magnification x40 at the bottom & D: In this double staining IL17 and CD3, the goat anti-human IL-17 antibody was added first before staining with Naphthol/Fast (red) followed by the rabbit anti-human CD3 antibody that was revealed with DAB substrate (brown); this showed that CD3 was not the only cell producing IL-17.

Article Snippet: For IL17/CD3 double staining, the goat anti-human IL-17 antibody (diluted 1∶40) was added for 2 h, and then staining was undertaken using Vectastain AP kit from Vector Laboratories (Burlingame, CA, USA), and revealed by Naphtol/Fast Red (Sigma-Aldrich).

Techniques: DNA Extraction, Double Staining, Staining