Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-17RA and IL-17RC receptors are essential for IL-17A-induced ELR+ CXC chemokine expression in synoviocytes and are overexpressed in rheumatoid blood.

doi: 10.4049/jimmunol.180.1.655

Figure Lengend Snippet: FIGURE 1. IL-17RA and IL-17RC mRNA expression in blood. A, IL-17RA and IL-17RC mRNA expression in peripheral whole blood from 40 RA patients (31 patients with severe RA and 9 with moderate RA) and from 19 healthy volunteers (HV), as deter- mined by microarray analysis. Results are presented as fluorescent intensity (, p 0.05; , p 0.005; , p 0.0005 by Mann-Whitney U test). B, IL-17RA or IL- 17RC were detected in whole blood from HV (n 3) or RA patients (n 6) by Western blotting using actin as a loading control. Den- sity ratios of IL-17RA or IL-17RC were nor- malized to actin and expressed as AU (, p 0.05 by Mann-Whitney test). C, IL-17RA and IL-17RC mRNA expression in peripheral blood subsets from three RA patients. CD14 monocytes, CD3 lymphocytes, and CD19 lymphocytes were isolated from PBMC by positive selection. IL-17RA and IL-17RC mRNA expression were assessed by real-time RT-PCR. PPIB was used to nor- malize gene expression. Values represent the mean SEM expression from three RA pa- tients (, p 0.05 by Mann-Whitney test).

Article Snippet: Human rIL-17A, the mAb against human IL-17RA and the polyclonal goat Ab against IL-17RC, were purchased from R&D Systems.

Techniques: Expressing, Microarray, MANN-WHITNEY, Western Blot, Control, Isolation, Selection, Quantitative RT-PCR, Gene Expression

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-17RA and IL-17RC receptors are essential for IL-17A-induced ELR+ CXC chemokine expression in synoviocytes and are overexpressed in rheumatoid blood.

doi: 10.4049/jimmunol.180.1.655

Figure Lengend Snippet: FIGURE 2. Detection of IL-17RA and IL-17RC in RA synovium. Im- munostaining of paraffin-embedded serial sections using mouse anti-IL- 17RA mAb (A and B, respectively, 200 and 400) and goat anti-IL- 17RC polyclonal Ab (C and D, respectively, 200 and 400) shows a diffuse positive staining (brown) in both the lining and sublining area. Control staining was performed using mouse IgG1 and goat serum (200) (insets in A and C). Immunostaining of IL-17RA and IL-17RC were also performed in OA synovium (E and F, respectively; 200). Some of the IL-17A-producing cells have a plasmacytoid-like appearance as suggested by the remote nucleus in a large cytoplasm (inset in D, 600).

Article Snippet: Human rIL-17A, the mAb against human IL-17RA and the polyclonal goat Ab against IL-17RC, were purchased from R&D Systems.

Techniques: Staining, Control, Immunostaining

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-17RA and IL-17RC receptors are essential for IL-17A-induced ELR+ CXC chemokine expression in synoviocytes and are overexpressed in rheumatoid blood.

doi: 10.4049/jimmunol.180.1.655

Figure Lengend Snippet: FIGURE 4. Effect of IL-17RA siRNA and IL-17RC siRNA on IL-17A- or IL-17F-induced IL-6 and IL-8 secretion by RA synoviocytes. RA synovio- cytes were transfected with IL-17RA siRNA and IL-17RC siRNA at 0.5 and 0.05 g, respectively. A scrambled nonsilence siRNA, siCONTROL, was used as a negative control. A, Knockdown efficiency of IL-17RA siRNA and IL-17RC siRNA was assessed by real-time RT-PCR. The values represent data obtained 24 h posttransfection and are expressed as mean SEM of three independent experiments (, p 0.05 by Dunnett’s test). B, Knockdown efficiency of IL-17RA siRNA and IL-17RC siRNA was assessed by Western blotting 48 h posttransfection, using actin as a loading control. A representative western blot among three separate experiments is shown. C, Effect of IL-17RA siRNA and IL-17RC siRNA on IL-17A-induced IL-6 and IL-8 secretion by RA synoviocytes. Forty-eight hours after siRNA delivery (siCONTROL, IL-17RA siRNA, or IL-17RC siRNA), RA synoviocytes were stimulated for 12 h with IL-17A at 50 ng/ml or TNF- at 0.5 ng/ml. IL-6 and IL-8 levels were quantified in removed supernatants by ELISA. The values are expressed as fold induction compared with the untreated condition (mean SEM in untreated cells transfected with siCONTROL siRNA: 2.4 0.8 ng/ml and 367 39.6 pg/ml, respectively) and are represented the mean SEM of three independent experiments (, p 0.05 by Dunnett’s test).

Article Snippet: Human rIL-17A, the mAb against human IL-17RA and the polyclonal goat Ab against IL-17RC, were purchased from R&D Systems.

Techniques: Transfection, Negative Control, Knockdown, Quantitative RT-PCR, Western Blot, Control, Enzyme-linked Immunosorbent Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-17RA and IL-17RC receptors are essential for IL-17A-induced ELR+ CXC chemokine expression in synoviocytes and are overexpressed in rheumatoid blood.

doi: 10.4049/jimmunol.180.1.655

Figure Lengend Snippet: FIGURE 5. Effect of IL-17RA and/or IL- 17RC inhibition by siRNA or extracellular inhib- itors, on IL-17 plus TNF--induced IL-6 secre- tion. A, Efficiency of posttranscriptional inhibition of IL-17RA and/or IL-17RC using siRNA. RA synoviocytes from the three cell pop- ulations (transfected with siCONTROL, IL- 17RA siRNA, and IL-17RC siRNA) were stim- ulation for 36 h with IL-17A (50 ng/ml), alone or in combination with TNF- (0.5 ng/ml). IL-6 levels were quantified in removed supernatants by ELISA. The values represent the mean SEM of three independent experiments (, p 0.05 by Dunnett’s test). B and C, Efficiency of extracellular inhibition using Abs. RA synovio- cytes were preincubated for 2 h (37°C, 5% CO2) with anti-IL-17RA Ab (10 g/ml) and/or anti- IL-17RC Ab (10 g/ml). The cells were then stimulated with IL-17A (50 ng/ml), alone or in combination with TNF- (0.5 ng/ml). IL-6 and CCL20 levels were quantified in removed super- natants by ELISA. The values represent the mean SEM of three independent experiments (mean SEM in untreated cells: 1.7 0.6 ng/ml and 1.4 1.6 pg/ml, respectively) (, p 0.05 by Dunnett’s test).

Article Snippet: Human rIL-17A, the mAb against human IL-17RA and the polyclonal goat Ab against IL-17RC, were purchased from R&D Systems.

Techniques: Inhibition, Transfection, Enzyme-linked Immunosorbent Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Th17-Derived Cytokines Synergistically Enhance IL-17C Production by the Colonic Epithelium.

doi: 10.4049/jimmunol.2200125

Figure Lengend Snippet: FIGURE 2. TNF-a stimulation does not prime cells to respond to IL-17A or IL-22. (A) Cells were stimulated with the stated combinations of cytokines for 3 h, and cells were prepared for flow cytometry analysis detecting IL-17RA, IL-17RC, IL-10Rb, and IL-22Ra1. (B) Cells were stimulated with the stated combinations of cytokines for 5 h, or IL-17A or IL-22 was added 3 h after stimulation with TNF-a, as noted. RNA was extracted and analyzed by RT-qPCR, where IL-17C fold induction (2−DDCt) was controlled against GAPDH mRNA levels. Statistical analysis was completed using a one-way ANOVA with a Dunnett’s multiple comparison test as shown. ****p < 0.0001. Each data point represents three separate experiments of biological replicates.

Article Snippet: Abs used were mouse anti-RelA (1:1000 dilution of F-6, Santa Cruz), mouse anti-p50 (1:500 dilution of E-10, Santa Cruz), mouse anti heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1; 1:5000 dilution of 4B10, Santa Cruz), mouse anti-GAPDH (1:1000 dilution of 6C5, Thermo Fisher Scientific), rabbit anti IL22Ra1 (1:1000 dilution of ab5984, Abcam), goat anti IL-17RA (1:1000 dilution of AF177, R&D Systems), rabbit anti-p38 (1:5000 dilution of D13E1, Cell Signaling Technology), rabbit anti phospho-p38 (1:1000 dilution of 9211, Cell Signaling Technology), rabbit anti-ERK1/2 (1:10,000 dilution of EPR17526, Abcam), rabbit anti phospho-ERK1/2 (1:500 dilution of 197G2, Abcam), rabbit anti-JNK1/2/3 (1:1000 dilution of EPR167979, Abcam), rabbit anti phosphoJNK1/2/3 (1:2000 dilution of EPR5693, Abcam), rabbit anti activating transcription factor (ATF)-2 (1:1000 dilution of D4L2X, Cell Signaling Technology), and rabbit anti phospho-ATF-2 (1:1000 dilution of A8J7P, Cell Signaling Technology).

Techniques: Flow Cytometry, Quantitative RT-PCR, Comparison

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Th17-Derived Cytokines Synergistically Enhance IL-17C Production by the Colonic Epithelium.

doi: 10.4049/jimmunol.2200125

Figure Lengend Snippet: FIGURE 7. Primary non-IBD and UC uninvolved human colonic spheroids, but not UC involved cells, stimulated with Th17-derived cytokines synergistically induce IL-17C expression. (A and B) Primary human colonic spheroids from nonIBD-affected individuals (A) and UC donors (B) were stimulated with TNF-a (50 ng/ml), IL-17A (100 ng/ml), and IL-22 (10 ng/ml). RNA was collected at 5 h, and IL-17C mRNA was quantified as previously described in the legend to Fig. 1. Statistical analysis was completed using a one-way ANOVA with a Dunnett’s multiple comparison test. **p 5 0.0035, ****p < 0.0001. Each data point repre- sents separate experiments of biological replicates for distinct spheroids derived from different donors. (C) Spheroids from non-IBD individuals and UC donors were prepared for flow cytometry analysis detecting IL-17RA, IL-17RC, IL-10Rb, and IL-22Ra1.

Article Snippet: Abs used were mouse anti-RelA (1:1000 dilution of F-6, Santa Cruz), mouse anti-p50 (1:500 dilution of E-10, Santa Cruz), mouse anti heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1; 1:5000 dilution of 4B10, Santa Cruz), mouse anti-GAPDH (1:1000 dilution of 6C5, Thermo Fisher Scientific), rabbit anti IL22Ra1 (1:1000 dilution of ab5984, Abcam), goat anti IL-17RA (1:1000 dilution of AF177, R&D Systems), rabbit anti-p38 (1:5000 dilution of D13E1, Cell Signaling Technology), rabbit anti phospho-p38 (1:1000 dilution of 9211, Cell Signaling Technology), rabbit anti-ERK1/2 (1:10,000 dilution of EPR17526, Abcam), rabbit anti phospho-ERK1/2 (1:500 dilution of 197G2, Abcam), rabbit anti-JNK1/2/3 (1:1000 dilution of EPR167979, Abcam), rabbit anti phosphoJNK1/2/3 (1:2000 dilution of EPR5693, Abcam), rabbit anti activating transcription factor (ATF)-2 (1:1000 dilution of D4L2X, Cell Signaling Technology), and rabbit anti phospho-ATF-2 (1:1000 dilution of A8J7P, Cell Signaling Technology).

Techniques: Derivative Assay, Expressing, Comparison, Flow Cytometry

Journal: Frontiers in Physiology

Article Title: IL-17A activates the PI3K/AKT/mTOR pathway to regulate bronchial fibroblast autophagy-mediated airway remodeling: evidence from conditional IL-17RA-deficient mice

doi: 10.3389/fphys.2026.1873777

Figure Lengend Snippet: Culture of primary bronchial fibroblasts with immunofluorescence evaluation of passaged MBFs. A, (B) Representative protein bands on WB with quantitative determination of IL-17RA in the different groups (n=5); (C) Primary bronchial fibroblasts on Days 3, 5, and 7; (D) Demonstration of vimentin expression in passaged MBFs by immunofluorescence. DAPI staining for double-stranded DNA. Data are presented as mean ± SD. Statistical analysis was performed using an unpaired Student’s t-test. ** P < 0.01 versus WT mice.

Article Snippet: Specific antibodies against autophagy proteins P62 (1:1000, #5114, CST), LC3II/I (1:1000, #4108, CST), type III collagen (1:1000, #A0817, ABclonal), and type I collagen (1:1000, #A21059, ABclonal), IL-17RA (1:1000, #A5163, ABclonal), and GAPDH (1:10000, #AC001, ABclonal) was used as a loading control.

Techniques: Immunofluorescence, Expressing, Staining

Journal: Biomedicines

Article Title: Defect of IL17 Signaling, but Not Centrinone, Inhibits the Development of Psoriasis and Skin Papilloma in Mouse Models

doi: 10.3390/biomedicines10081976

Figure Lengend Snippet: Expression pattern of IL17 and PLK family members in skin cancer and autoimmune diseases. ( A , B ) Original data of GSE109248 were analyzed using the OmicSoft platform. The heatmap shows the expression patterns of PLK family members among normal skin, cutaneous lupus erythematosus, and psoriasis. ( C ) The GEO2R tool was applied to differentially determine the expressed genes ( p value ≤ 0.01) between psoriasis and normal skin. ( D ) The heatmap shows expression patterns of IL17 family members and PLK family members among normal skin, actinic keratosis (AK), and squamous cell carcinoma (SCC) of skin. Data were downloaded from GDS2200 database. ( E ) The volcano plot demonstrates significant genes between SCC and normal skin. Data were downloaded from GSE2503. ( F ) Immunohistochemical staining of human tissues from The Human Protein Atlas (HPA, https://www.proteinatlas.org/ , accessed on 12 July 2022) shows expression patterns of IL17RA, IL17RC, and PLK4 between normal skin and SCC.

Article Snippet: Immunohistochemical staining of human tissues from the HPA database were downloaded to show the expression pattern of IL17RA (cat# MAB1771, R&D system, Minneapolis, MN, USA), IL17RC (cat# AF2269, R&D system, Minneapolis, MN, USA) and PLK4 (cat# HPA035026, Millipore Sigma, St. Louis, MO, USA) between normal skin and SCC.

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Biomedicines

Article Title: Defect of IL17 Signaling, but Not Centrinone, Inhibits the Development of Psoriasis and Skin Papilloma in Mouse Models

doi: 10.3390/biomedicines10081976

Figure Lengend Snippet: Effect of IL17RA knockdown and centrinone on squamous skin cancer cells. ( A ) Endogenous IL17RA of HaCaT and A431 was knocked down using small interference RNA of IL17RA. ( B , C ) Colony formation assay was conducted to determine the effect of IL17RA knockdown and inhibition of PLK4 on the proliferation of normal skin cell line HaCaT and squamous skin cancer cell line A431. A two-tailed Student’s t test was applied to analyze statistical significance. Error bar represents mean ± standard deviation (SD); ns—no significance; ** — p < 0.01.

Article Snippet: Immunohistochemical staining of human tissues from the HPA database were downloaded to show the expression pattern of IL17RA (cat# MAB1771, R&D system, Minneapolis, MN, USA), IL17RC (cat# AF2269, R&D system, Minneapolis, MN, USA) and PLK4 (cat# HPA035026, Millipore Sigma, St. Louis, MO, USA) between normal skin and SCC.

Techniques: Knockdown, Colony Assay, Inhibition, Two Tailed Test, Standard Deviation

Journal: Biomedicines

Article Title: Defect of IL17 Signaling, but Not Centrinone, Inhibits the Development of Psoriasis and Skin Papilloma in Mouse Models

doi: 10.3390/biomedicines10081976

Figure Lengend Snippet: Knockout of Il17rc alleviates skin lesions induced by imiquimod. ( A ) Diagram of establishing the imiquimod-induced psoriasis mouse model and skin lesions of WT C57BL/6J, Il17ra (T779A)-knockin, and Il17rc -knockout strains. ( B ) Representatives of hematoxylin and eosin stain of skin lesions. Scale bar represents 100 µm. ( C ) To analyze the statistical significance of the thickness of epidermis, a two-tailed Student’s t test was applied. Error bar represents mean ± SD. **** indicates p < 0.0001. The numbers of male and female mice in each group are indicated in the panel.

Article Snippet: Immunohistochemical staining of human tissues from the HPA database were downloaded to show the expression pattern of IL17RA (cat# MAB1771, R&D system, Minneapolis, MN, USA), IL17RC (cat# AF2269, R&D system, Minneapolis, MN, USA) and PLK4 (cat# HPA035026, Millipore Sigma, St. Louis, MO, USA) between normal skin and SCC.

Techniques: Knock-Out, Knock-In, H&E Stain, Two Tailed Test

Journal: Biomedicines

Article Title: Defect of IL17 Signaling, but Not Centrinone, Inhibits the Development of Psoriasis and Skin Papilloma in Mouse Models

doi: 10.3390/biomedicines10081976

Figure Lengend Snippet: Mutation of T779A of Il17ra and knockout of Il17rc prevent the development of skin papilloma. ( A ) Diagram of establishing DMBA/TPA-induced two-stage skin papilloma mouse model. ( B ) Representative of tumors or nodules of WT, Il17ra (T779A)-knockin, and Il17rc -knockout C57BL/6J mouse strains. Arrowheads indicate papillomata (≥1mm) and nodules (<1 mm). ( C – E ) Plots demonstrate tumor incidence, tumor multiplicity, and tumor volume. Statistical significance was assessed with a two-tailed Student’s t test and two-way ANOVA. Error bar represents mean ± SD. *— p < 0.05; **— p < 0.01. The number of male and female mice in each subgroup are indicated in the panel.

Article Snippet: Immunohistochemical staining of human tissues from the HPA database were downloaded to show the expression pattern of IL17RA (cat# MAB1771, R&D system, Minneapolis, MN, USA), IL17RC (cat# AF2269, R&D system, Minneapolis, MN, USA) and PLK4 (cat# HPA035026, Millipore Sigma, St. Louis, MO, USA) between normal skin and SCC.

Techniques: Mutagenesis, Knock-Out, Knock-In, Two Tailed Test