Journal: Scientific Reports

Article Title: CD4+ and γδ T Cells are the main Producers of IL-22 and IL-17A in Lymphocytes from Mycobacterium bovis -infected Cattle

doi: 10.1038/srep29990

Figure Lengend Snippet: Summary of flow cytometry antibody labelling panels a .

Article Snippet: CD4+ T cell population , 1 ° : anti-boCD4-AF647 , 1 ° : boil-22 Fab antibody (clone 58, Bio-Rad AbD Serotec), polyclonal rabbit anti-boIL-17A-biotin (Kingfisher Biotech)2 ° : anti-huIgG F(ab’) 2 -FITC, Streptavidin-PE (BD Pharmingen).

Techniques: Flow Cytometry, Clone Assay, Recombinant

Journal: PLoS ONE

Article Title: Bone marrow derived-mesenchymal stem cells downregulate IL17A dependent IL6/STAT3 signaling pathway in CCl 4 -induced rat liver fibrosis

doi: 10.1371/journal.pone.0206130

Figure Lengend Snippet: Sequence of primers used in RT-qPCR analysis.

Article Snippet: Slides were washed with TBS-Triton three times and one time with TBS for 5 min. Nonspecific binding was blocked using 5% goat serum for 30 min and the blocking buffer was then removed and sections were incubated with a polyclonal anti–IL17A antibody (1:2000) (USBiological, Salem, MA, USA), polyclonal anti–IL17RA antibody (2.5μg/ml) (GeneTex, Irvine, CA, USA) and polyclonal antibody to PCNA (2.5mg/ml) (ThermoFisher Scientific, Rockford, IL, USA).

Techniques: Sequencing

Journal: PLoS ONE

Article Title: Bone marrow derived-mesenchymal stem cells downregulate IL17A dependent IL6/STAT3 signaling pathway in CCl 4 -induced rat liver fibrosis

doi: 10.1371/journal.pone.0206130

Figure Lengend Snippet: The data are presented as fold induction of mRNA expression of a . Il17a , b . Il17f , c . Il17ra , and d . Il17rc in CCl 4 /untreated (n = 5) rats compared to MSCs transplanted (CCl 4 +BM-MSCs) groups (n = 5) after 2, 4 and 6 weeks. All groups are compared with normal/diet control and oil/vehicle control rats (n = 5). Each experiment was performed in duplicates. Representative data are shown as mean±SD. * P <0.05. # P <0.05 compared to control groups.

Article Snippet: Slides were washed with TBS-Triton three times and one time with TBS for 5 min. Nonspecific binding was blocked using 5% goat serum for 30 min and the blocking buffer was then removed and sections were incubated with a polyclonal anti–IL17A antibody (1:2000) (USBiological, Salem, MA, USA), polyclonal anti–IL17RA antibody (2.5μg/ml) (GeneTex, Irvine, CA, USA) and polyclonal antibody to PCNA (2.5mg/ml) (ThermoFisher Scientific, Rockford, IL, USA).

Techniques: Expressing

Journal: PLoS ONE

Article Title: Bone marrow derived-mesenchymal stem cells downregulate IL17A dependent IL6/STAT3 signaling pathway in CCl 4 -induced rat liver fibrosis

doi: 10.1371/journal.pone.0206130

Figure Lengend Snippet: Upper panels are representatives of IL17 expression and it is noted that it was very weak in normal and paraffin/oil control group, severe in CCl 4 /fibrosis and recovery groups, high in CCl 4 +BM-MSCs/2 weeks group, moderate in CCl 4 +BM-MSCs/4weeks group and mild CCl 4 +BM-MSCs/6 weeks group. Lower panels are representatives of IL17RA expression and it is noted that it was absent in normal and paraffin/oil control group, high in CCl 4 /fibrosis and recovery groups, moderate in CCl 4 +BM-MSCs/2weeks group, mild in CCl 4 +BM-MSCs/4weeks group and nil in CCl 4 +BM-MSCs/6 weeks group (400X).

Article Snippet: Slides were washed with TBS-Triton three times and one time with TBS for 5 min. Nonspecific binding was blocked using 5% goat serum for 30 min and the blocking buffer was then removed and sections were incubated with a polyclonal anti–IL17A antibody (1:2000) (USBiological, Salem, MA, USA), polyclonal anti–IL17RA antibody (2.5μg/ml) (GeneTex, Irvine, CA, USA) and polyclonal antibody to PCNA (2.5mg/ml) (ThermoFisher Scientific, Rockford, IL, USA).

Techniques: Expressing

Journal: Oncoimmunology

Article Title: Th17 cell-derived IL-17A promoted tumor progression via STAT3/NF-κB/Notch1 signaling in non-small cell lung cancer

doi: 10.1080/2162402X.2018.1461303

Figure Lengend Snippet: The level of Th17 cells in NSCLC was increased. (A) CD3+CD4+IL-17░A+IFN-γ−Th17 cells from peripheral blood were analyzed by flow cytometry in NSCLC patients and healthy donors. (B) Comparison of Th17 cell phenotypes in peripheral blood from NSCLC patients and healthy donors represented as scatter. (C) Comparison of Th17 cell frequencies in NSCLC tissues and non-tumor tissues represented as scatter. (D) Comparison of Th17 cell frequencies in peripheral blood and tumor tissues from NSCLC patients represented as scatter. (E) Tumor tissues and non-tumor tissues from NSCLC patients were subjected to double immunofluorescence for CD4 (red), IL-17░A (green) and DAPI (blue). *, ** and *** indicate P < 0.05, P < 0.01 and P< 0.001, respectively. Scale bar represents 50 μm.

Article Snippet: Mouse anti-CD4 (Abcam, #ab846), rabbit anti-IL-17░A (PeproTech, #500-P07) and mouse anti-CD31 (R&D, #BBA7) were used as the primary antibodies.

Techniques: Flow Cytometry, Immunofluorescence

Journal: Oncoimmunology

Article Title: Th17 cell-derived IL-17A promoted tumor progression via STAT3/NF-κB/Notch1 signaling in non-small cell lung cancer

doi: 10.1080/2162402X.2018.1461303

Figure Lengend Snippet: IL-17░A promoted the migration and invasion of NSCLC cells in vitro. The migration activities of A549 (A) and H460 (B) cells, and the invasion activities of A549 (C) and H460 (D) cells before and after treatment of rhIL-17░A was analyzed using transwell assay. One representative analysis is shown. Data are presented as a histogram. (E) The expression of N-cadherin in A549 and H460 cells before and after treatment of rhIL-17░A was analyzed using western blotting. (F) The relative expression of N-cadherin in A549 and H460 cells before and after treatment with rhIL-17░A was analyzed using qPCR. (G) The expressions of IL-17░A and CD31 in tumor tissues from NSCLC patients were detected by immunofluorescence. * indicates P < 0.05. Scale bar represents 50 μm.

Article Snippet: Mouse anti-CD4 (Abcam, #ab846), rabbit anti-IL-17░A (PeproTech, #500-P07) and mouse anti-CD31 (R&D, #BBA7) were used as the primary antibodies.

Techniques: Migration, In Vitro, Transwell Assay, Expressing, Western Blot, Immunofluorescence

Journal: Oncoimmunology

Article Title: Th17 cell-derived IL-17A promoted tumor progression via STAT3/NF-κB/Notch1 signaling in non-small cell lung cancer

doi: 10.1080/2162402X.2018.1461303

Figure Lengend Snippet: The STAT3/NF-κB/Notch1 signaling was critical for IL-17░A-induced migration and invasion in NSCLC cells. (A) The activation of STAT3, NF-κB and Notch1in A549 and H460 cells treated with rhIL-17░A was analyzed using western blotting. (B) The migration activities of A549 and H460 cells treated with or without rhIL-17░A and STAT3, or NF-κB, or Notch1 inhibitor were assessed by transwell assay. One representative analysis is shown. The data from A549 (C) and H460 (D) cells are presented as histogram. (E) The invasion activities of A549 and H460 cells treated with or without rhIL-17░A and these molecular inhibitors were assessed by transwell assay. One representative analysis is shown. The data from A549 (F) and H460 (G) cells are presented as a histogram. (H) The expression of N-cadherin in A549 and H460 cells treated with or without rhIL-17░A and STAT3, or NF-κB, or Notch1 inhibitor was analyzed using western blotting. * indicates P < 0.05. Scale bar represents 50 μm.

Article Snippet: Mouse anti-CD4 (Abcam, #ab846), rabbit anti-IL-17░A (PeproTech, #500-P07) and mouse anti-CD31 (R&D, #BBA7) were used as the primary antibodies.

Techniques: Migration, Activation Assay, Western Blot, Transwell Assay, Expressing

Journal: Oncoimmunology

Article Title: Th17 cell-derived IL-17A promoted tumor progression via STAT3/NF-κB/Notch1 signaling in non-small cell lung cancer

doi: 10.1080/2162402X.2018.1461303

Figure Lengend Snippet: IL-17░A promoted the CSC-like properties of NSCLC cells. A549 (A) and H460 (B) cells were cultured with rhIL-17░A for 7 days, and then collected for sphere assay. One representative photomicrograph is shown. Data are presented as a histogram. (C) The expression of Oct4 in A549 and H460 cells before and after treatment of rhIL-17░A was analyzed using western blotting. The apoptosis of A549 (D) and H460 (E) cells treated with or without docetaxel and rhIL-17░A was analyzed by flow cytometry. IL-17░A expression in IL-17░A-expressing and control vectors-tranfected H460 cells was analyzed by western blotting (F) and qPCR (G). (H) IL-17░A promotes NSCLC tumor growth in vivo. IL-17░A expressing A549 cells were injected subcutaneously into nude mice (n = 6 mice/group). Tumor volumes were measured at 21 days after cell implantation. (I) Lump images of xenograft tumors that were formed in nude mice at 21 days after cell implantation. (J) The result of tumor weights at 21 days after cell implantation was shown as histogram. (K) Tumor growth curve from 7 to 21 days after cell implantation was shown as statistical graph. * indicates P < 0.05. Scale bar represents 100 μm.

Article Snippet: Mouse anti-CD4 (Abcam, #ab846), rabbit anti-IL-17░A (PeproTech, #500-P07) and mouse anti-CD31 (R&D, #BBA7) were used as the primary antibodies.

Techniques: Cell Culture, Expressing, Western Blot, Flow Cytometry, In Vivo, Injection

Journal: Oncoimmunology

Article Title: Th17 cell-derived IL-17A promoted tumor progression via STAT3/NF-κB/Notch1 signaling in non-small cell lung cancer

doi: 10.1080/2162402X.2018.1461303

Figure Lengend Snippet: Blockade of STAT3/NF-κB/Notch1 signaling inhibited NSCLC stemness promoted by IL-17░A. (A) After the treatment with STAT3░or NF-κB or Notch1 inhibitor, the sphere forming of A549 and H460 cells induce by rhIL-17░A was blocked. One representative photomicrograph is shown. The results from A549 (B) and H460 (C) cells were shown as histogram. (D) The expression of Oct4 in A549 and H460 cells treated with or without rhIL-17░A and these molecular inhibitors were assessed by western blotting. The apoptosis of A549 (E) and H460 (F) cells treated with or without rhIL-17░A and these molecular inhibitors were assessed by transwell assay. * indicates P < 0.05. Scale bar represents 100 μm.

Article Snippet: Mouse anti-CD4 (Abcam, #ab846), rabbit anti-IL-17░A (PeproTech, #500-P07) and mouse anti-CD31 (R&D, #BBA7) were used as the primary antibodies.

Techniques: Expressing, Western Blot, Transwell Assay

Journal: Oncoimmunology

Article Title: Th17 cell-derived IL-17A promoted tumor progression via STAT3/NF-κB/Notch1 signaling in non-small cell lung cancer

doi: 10.1080/2162402X.2018.1461303

Figure Lengend Snippet: The level of Th17 cells was associated with poor survival in NSCLC patients. The relationship between Th17 frequency and TNM stage (A), tumor grade (B) and lymph node metastasis (C) of NSCLC patients, respectively. The changes of Th17 cell frequency in peripheral blood from NSCLC patients before and after chemotherapy with the status of PD (D), PR (E) and SD (F). (G) IL-17░A expression in tumor tissues of NSCLC was analyzed by immunohistochemistry. (H) Kaplan-Meier survival curves for NSCLC patients with lower and higher IL-17░A expression (immunohistochemistry analysis). *, ** and ns indicate P < 0.05, P < 0.05 and non-significance, respectively. Scale bar represents 50 μm.

Article Snippet: Mouse anti-CD4 (Abcam, #ab846), rabbit anti-IL-17░A (PeproTech, #500-P07) and mouse anti-CD31 (R&D, #BBA7) were used as the primary antibodies.

Techniques: Expressing, Immunohistochemistry