Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Interleukin-17 Cytokines and Receptors: Potential Amplifiers of Tendon Inflammation

doi: 10.3389/fbioe.2021.795830

Figure Lengend Snippet: Representative images showing protein expression of IL-17A and IL-17 receptor A (IL-17RA) and IL-17RC in healthy tendon tissue (hamstring tendon), or diseased tendon tissue ranging from tendinopathic (untorn) to small, medium, large, and massive rotator cuff tears (supraspinatus tendon). Antibodies were visualised with DAB (brown) and nuclei counterstained with haematoxylin (blue). Images were taken at 40x magnification; scale bar = 100 μm.

Article Snippet: Immunostaining was performed using an Autostainer Link 48 machine using the EnVision FLEX visualisation system (Dako) with anti-human IL-17RA, anti-human IL-17RC antibodies (R&D systems, Abingdon, United Kingdom) or universal negative control mouse (Dako) ( ).

Techniques: Expressing

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Interleukin-17 Cytokines and Receptors: Potential Amplifiers of Tendon Inflammation

doi: 10.3389/fbioe.2021.795830

Figure Lengend Snippet: Representative images of the protein expression of podoplanin (PDPN, green) and IL-17RA or IL-17RC (red) in supraspinatus tendons with a large/massive tear. POPO-1 staining was used to visualise nuclei (cyan). Images taken at 40x; scale bar = 20 μm.

Article Snippet: Immunostaining was performed using an Autostainer Link 48 machine using the EnVision FLEX visualisation system (Dako) with anti-human IL-17RA, anti-human IL-17RC antibodies (R&D systems, Abingdon, United Kingdom) or universal negative control mouse (Dako) ( ).

Techniques: Expressing, Staining

Journal: Respiratory research

Article Title: Increased IL-17RA and IL-17RC in End-Stage COPD and the Contribution to Mast Cell Secretion of FGF-2 and VEGF.

doi: 10.1186/s12931-017-0534-9

Figure Lengend Snippet: Fig. 1 Elevated expression of IL-17RA and IL-17RC in end-stage COPD. Bright field micrographs of (a) interleukin (IL)-17 receptor (R)A in never smoker, (b) IL-17RA in COPD, (c) IL-17RC in never smoking non-COPD controls and (d) IL-17RC in COPD. Number of immunopositive cells to (e) interleukin (IL)-17RA and (f) IL-17RC in lung tissue of never smokers, smokers and patients with GOLD I-II or III-IV COPD. Horizontal line indicates mean value. One-way ANOVA and Dunnet’s test (GraphPad Prism 6, GraphPad Software, La Jolla, CA) was used to detect differences between GOLD IV COPD and all other groups. Clear circles: never smokers; grey circles: asymptomatic smokers, triangles (up): GOLD I COPD; triangles (down): GOLD II COPD; squares: GOLD III; diamonds: GOLD IV COPD. n = 7-18. *p < 0.05, **p < 0.01, *p < 0.001 compared to GOLD IV COPD. Co-immunohistochemical staining for (g-k) interleukin (IL)-17RA and (l-p) IL-17RC (DAB, brown) and cellular markers (Vina green, blue-green). Sections were counterstained with Mayer’s Hematoxylin. Arrows in indicate double positive cells. Scale bar: 100 μm. Expression of (q) FGF-2, and (r) VEGF in cell supernatant of stem cell-derived mast cells stimulated with 1000 ng/ml IL-17A 8 hours. Experiments were performed in triplicates and repeated twice. Bars indicate mean value, errorbars indicate SEM. n = 5. *p < 0.05. Light micrograph of (s) fibroblast growth factor (FGF)-2 (DAB; brown) and (t) vascular endothelial growth factor (VEGF) (DAB; brown) in GOLD IV COPD. Immunoreactivity was detected by DAB (brown) and sections were counterstained with Mayer’s Hematoxylin (blue). Mast cells were identified by immunodetection of tryptase (Vena green; green-blue). Arrows indicate FGF-2 and VEGF positive mast cells. Scale bar indicates 100 μm

Article Snippet: We used automated immunohistochemistry to detect the expression of IL-17RA (mouse anti-IL17RA, R&D Systems, Minneapolis, MN, USA) and IL-17RC (mouse anti-IL17RC, Atlas Antibodies, Stockholm, Sweden) in peripheral lung tissue sections obtained from a previously characterized cohort of COPD patients and relevant controls (Table 1) [3].

Techniques: Expressing, Software, Immunohistochemical staining, Staining, Derivative Assay, Immunodetection

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Interleukin-17 signaling mediates cytolytic natural killer cell activation in response to placental ischemia

doi: 10.1152/ajpregu.00285.2019

Figure Lengend Snippet: Effects of chronic IL-17 inhibition on litter size and fetal and placental weight in pregnant rats

Article Snippet: A subset of rats undergoing the RUPP procedure also received a miniosmotic pump (Alzet model 2002, Alzet Scientific, Cupertino, CA) that infused 100 pg/day of recombinant mouse IL-17 receptor C (IL-17RC) (R&D Systems Minneapolis, MN) into the intraperitoneal cavity from GD14–19.

Techniques: Inhibition

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Interleukin-17 signaling mediates cytolytic natural killer cell activation in response to placental ischemia

doi: 10.1152/ajpregu.00285.2019

Figure Lengend Snippet: Effects of chronic IL-17 inhibition on circulating and placental cytokines and cytolytic proteins in pregnant rats

Article Snippet: A subset of rats undergoing the RUPP procedure also received a miniosmotic pump (Alzet model 2002, Alzet Scientific, Cupertino, CA) that infused 100 pg/day of recombinant mouse IL-17 receptor C (IL-17RC) (R&D Systems Minneapolis, MN) into the intraperitoneal cavity from GD14–19.

Techniques: Inhibition

Journal: Cytokine

Article Title: When recombinant proteins go wrong: The hidden pitfall of recombinant protein contamination.

doi: 10.1016/j.cyto.2024.156830

Figure Lengend Snippet: Fig. 2. Confirmation of IL-4 Contamination in Recombinant IL-17F (PeproTech). (A) Cultured human fibroblasts were stimulated with IL-17A, IL-17F from both PeproTech and R&D Systems, and IL-4 (positive control) for 24 h to compare gene expression via qPCR (n = 6). The analysis initially aimed to compare IL-17A and IL- 17F, but unexpectedly, IL-17F from PeproTech induced IL-4-responsive genes (IL4R and CCL26). (B) Further tests with IL-17F from R&D Systems confirmed that pure IL-17F does not induce IL-4 genes. Data are presented as fold changes, normalized to RPLP0. Statistical significance: *p < 0.05, **p ≤0.01, ***p ≤0.001, ***p ≤ 0.0001, ns (not significant). (C) Cultured human fibroblasts were stimulated with recombinant IL-17F from PeproTech and R&D Systems for 15 min (n = 4). Western blot analysis shows STAT6 phosphorylation in cells treated with IL-17F from PeproTech, but not from R&D Systems. β-actin was used as the loading control. (D) Western blot analysis was performed to assess the presence of IL-4 in Recombinant Human IL-17F (Catalog# 200–25, Lot# 0312277) from PeproTech, purchased at two different time points. IL-4 was detected in the PeproTech IL-17F protein, while no IL-4 was present in the IL-17F from R&D Systems. These results confirm IL-4 contamination in the PeproTech recombinant IL-17F, which may misdescribe prior experimental findings attributed to IL-17F. For analysis, 0.5 μg of recombinant IL- 4 (PeproTech) was used as a positive control, along with 1 μg of Recombinant Human IL-17F from both PeproTech and R&D Systems. (E) Fibroblasts were stimulated with IL-17F (PeproTech) for 15 min. Antibodies against IL-17RA and IL-17RC were used to block IL-17 signaling. Mouse IgG1 kappa isotype served as a control. β-actin was the loading control.

Article Snippet: For experiments involving IL-17 receptor blockade, cultured fibroblasts were pre-incubated with antibodies against human IL-17RA receptor: Anti-hIL17RA (10 μg/mL; R&D Systems) and human IL- 17RC receptor: Anti-hIL17RC (10 μg/mL; R&D Systems).

Techniques: Recombinant, Cell Culture, Positive Control, Gene Expression, Western Blot, Phospho-proteomics, Control, Blocking Assay

Journal: Immunology and cell biology

Article Title: Follistatin-like protein 1 modulates IL-17 signaling via IL-17RC regulation in stromal cells

doi: 10.1038/icb.2017.26

Figure Lengend Snippet: Il17rc transcript abundance is reduced in (A) ST2 cells with shRNA FSTL-1 suppression as well as (B) primary BMSCs from FSTL-1 KO mice when compared to controls. Surface expression of IL-17RC is reduced as measured by FACS determined by (C) %IL17RC+ and (D) mean fluorescence intensity as noted by (E) representative histograms of isotype (dark grey), FSTL-1 KO (light grey) and WT BMSCs (dashed line). *p<0.05, **p<0.01.

Article Snippet: Murine Il17rc (Addgene 46864), Fstl1 (Origene MR204305) and empty vector pCMV6-Entry control (Origene PS 100001) plasmids were transformed in and grown in Mix and Go E. coli (Zymo Research) in 5ml Luria-Bertani broth containing selection antibiotic and prepared using the QIAprep Spin Miniprep kit (Qiagen 27106), per manufacturer’s instructions.

Techniques: shRNA, Expressing, Fluorescence

Journal: Immunology and cell biology

Article Title: Follistatin-like protein 1 modulates IL-17 signaling via IL-17RC regulation in stromal cells

doi: 10.1038/icb.2017.26

Figure Lengend Snippet: FSTL-1 KO BMSC transfection with pCMV- il17rc rescues Il17rc transcript abundance (A). pCMV- Il17rc complementation in FSTL-1 KO BMSCs rescues IL-17A stimulated transcript levels of (B) Il6 and (C) Csf3 . * p<0.05, ** p<0.01, ** p<0.001

Article Snippet: Murine Il17rc (Addgene 46864), Fstl1 (Origene MR204305) and empty vector pCMV6-Entry control (Origene PS 100001) plasmids were transformed in and grown in Mix and Go E. coli (Zymo Research) in 5ml Luria-Bertani broth containing selection antibiotic and prepared using the QIAprep Spin Miniprep kit (Qiagen 27106), per manufacturer’s instructions.

Techniques: Transfection

Journal: Immunology and cell biology

Article Title: Follistatin-like protein 1 modulates IL-17 signaling via IL-17RC regulation in stromal cells

doi: 10.1038/icb.2017.26

Figure Lengend Snippet: FSTL-1 KO BMSC transfection with pCMV- fstl1 rescues (A) Fstl1 and (B) Il17rc transcript abundance. pCMV- Fstl1 complementation in FSTL-1 KO BMSCs rescues IL-17A/TNF stimulated transcript levels of (C) Il6 and (D) Csf3 . *p<0.05, ** p<0.01

Article Snippet: Murine Il17rc (Addgene 46864), Fstl1 (Origene MR204305) and empty vector pCMV6-Entry control (Origene PS 100001) plasmids were transformed in and grown in Mix and Go E. coli (Zymo Research) in 5ml Luria-Bertani broth containing selection antibiotic and prepared using the QIAprep Spin Miniprep kit (Qiagen 27106), per manufacturer’s instructions.

Techniques: Transfection

Journal: Immunology and cell biology

Article Title: Follistatin-like protein 1 modulates IL-17 signaling via IL-17RC regulation in stromal cells

doi: 10.1038/icb.2017.26

Figure Lengend Snippet: Following ActinomycinD treatment, FSTL-1 KO cells had reduced (A) Fstl1 and (8) Il17rc transcript abundance before ActinomycinD treatment and at 1, 2, 4 and 6 hours post-treatment compared with WT BMSCs. Linear regression of Il17rc transcript abundance (C) at various timepoints following ActinomycinD treatment showed similar slopes for WT and FSTL-1 KO BMSCs (−3.254±1.200 and −2.457±1.429, p=0.6748). Newly synthesized mRNA (5-EU labeled) was similar for (D) Hprt , but reduced for (E) Fstl1 and (F) Il17rc , transcripts in FSTL-1 KO cells compared to WT * p<0.05, ** p<0.01, ***p<0.001, ****p<0.0001

Article Snippet: Murine Il17rc (Addgene 46864), Fstl1 (Origene MR204305) and empty vector pCMV6-Entry control (Origene PS 100001) plasmids were transformed in and grown in Mix and Go E. coli (Zymo Research) in 5ml Luria-Bertani broth containing selection antibiotic and prepared using the QIAprep Spin Miniprep kit (Qiagen 27106), per manufacturer’s instructions.

Techniques: Synthesized, Labeling