anti-human il-17c antibody  (Novus Biologicals)

Journal: JCI Insight

Article Title: Suppression of TCF4 promotes a ZC3H12A-mediated self-sustaining inflammatory feedback cycle involving IL-17RA/IL-17RE epidermal signaling

doi: 10.1172/jci.insight.172764

Figure Lengend Snippet: ( A ) Silencing TCF4 ( siRNA-mediated ) in N/TERT keratinocytes, but not NFKB1 , JUN , or CEBPG , increases IL17C mRNA expression, which is further increased in the presence of TNF-α stimulation (10 ng/mL; n = 3, mean ± SEM, 2-way ANOVA with post hoc Tukey test. * P < 0.01; ** P < 0.005, *** P < 0.001, **** P < 0.0001; dashed line and the diamond demarcate P < 0.05 between 2 indicated groups via Student’s t test . ( B ) ATAC-Seq of human KCs isolated from fresh tissue biopsies identifies TCF4 binding sites in open chromatin regions of IL17C and ZC3H12A promoters. ( C ) Representative images of healthy normal and of lesional and nonlesional Ps skin demonstrates decreases in TCF4 (nuclear localization) and increases in IL-17C and ZC3H12A staining (using IHC; stained protein appears brown in color). Insets represent higher-magnification image. Scale bar: 100 μm; 20 μm (insets). ( D ) IL17C expression negatively correlates with TCF4 and positively correlates with ZC3H12A in lesional Ps and AD skin.

Article Snippet: Slides were treated with 3% H 2 O 2 (5 minutes), blocked with 10% goat serum (30 minutes), and incubated with primary antibodies against TCF4 (HPA025958-100UL, Sigma-Aldrich), IL-17C (AF1234, R&D Systems), and rabbit IgG isotype control (NI01, Sigma-Aldrich) overnight at 4°C.

Techniques: Expressing, Isolation, Binding Assay, Staining

Journal: JCI Insight

Article Title: Suppression of TCF4 promotes a ZC3H12A-mediated self-sustaining inflammatory feedback cycle involving IL-17RA/IL-17RE epidermal signaling

doi: 10.1172/jci.insight.172764

Figure Lengend Snippet: ( A ) TCF4 decreases in KCs stimulated with IL-17C (200 ng/mL), IL-17A (20 ng/mL), TNF-α (10 ng/mL; 8 hours), and combinations of these cytokines as indicated. ( B ) Venn diagrams show the number of DEGs altered by IL-17C and IL-17A in keratinocytes with or without TNF-α stimulation. ( C ) GO BP terms. The chart shows functional categories enriched in keratinocytes induced by IL-17C and IL-17A. ( D ) Partial heatmap showing NFKBIZ and ZC3H12A increase in an IL-17C–IL17RA/RE–dependent manner. Full heatmap is presented in . ( E ) Top, TCF4 decreases in KCs engineered to have no IL17RE and IL17RA . Bottom, since ZC3H12A and NFKB had also been identified in the IL-17C promotor analyses , we focused on these target genes. Normalized counts of TCF4 , NFKBIZ, and ZC3H12A , following IL-17C stimulation in WT and IL17RA- , IL17RC- , and IL17RE -KO keratinocytes. ( F ) TCF4 decreases in KCs stimulated with TNF-α, IL-17A, and IL-17A + TNF-α and are dependent on IL-17RE/RA expression. n = 3, mean ± SEM, 1-way ANOVA ( A and E inset) and 2-way ANOVA ( E and F ) with post hoc Tukey test. * P <0.05, ** P < 0.002, *** P < 0.0005, **** P < 0.0001.

Article Snippet: Slides were treated with 3% H 2 O 2 (5 minutes), blocked with 10% goat serum (30 minutes), and incubated with primary antibodies against TCF4 (HPA025958-100UL, Sigma-Aldrich), IL-17C (AF1234, R&D Systems), and rabbit IgG isotype control (NI01, Sigma-Aldrich) overnight at 4°C.

Techniques: Functional Assay, Expressing

Journal: JCI Insight

Article Title: Suppression of TCF4 promotes a ZC3H12A-mediated self-sustaining inflammatory feedback cycle involving IL-17RA/IL-17RE epidermal signaling

doi: 10.1172/jci.insight.172764

Figure Lengend Snippet: Skin inflammation occurs in an IL-17C–IL-17RA/RE–dependent manner and negatively correlates with Tcf4 expression and positively correlates with Zc3h12a expression. ( A ) Images of representative KC-Tie2 mice and KC-Tie2 mice deficient in Il17c , Il17ra , and Il17re . Representative images of H&E-stained skin isolated from the demarcated region of each mouse and adjacent skin stained for CD4 + T cells. ( B ) Dot plots of epidermal thickness measures for each mouse (control, n = 14; KC-Tie2, n = 22; KC-Tie2 × Il17re –/– , n = 6; KC-Tie2 × Il17c –/– , n = 16; KC-Tie2 × Il17ra –/– , n = 4) within each mouse strain. ( C ) Dot plots for individual quantification of CD4 + T cell numbers/field of view in skin of each mouse line ( n = 3–9/group [grp]). ( D and E ) qPCR measures for Il17a ( D , n = 4–10/grp) and ELISA of IL-17A protein ( E , n = 4–6/grp) expression for individual mice within each group. ( F ) IL-17C protein expression (using ELISA) in skin of psoriasis mouse models ( n = 8/grp), including imiquimod, KC-Tie2, Klk6 + , and IL-17C + . ( G ) IL-17C protein expression (using ELISA) for KC-Tie2 mice and KC-Tie2 mice deficient in Il17c , Il17ra , and Il17re ( n = 4–7/grp). ( H and I ) qPCR of Tcf4 ( H ) and Zc3h12a ( I ) gene expression in each mouse line ( n = 4–7/grp). ( J ) IHC of TCF4 protein in skin shows decreases in nuclear staining in KC-Tie2 mice compared with staining in control (WT) mice and KC-Tie2 mice deficient in Il17c , Il17ra , and Il17re . Insets represents higher-magnification image. One-way ANOVA, followed by post hoc Tukey test; * P < 0.05; ** P < 0.002; *** P <0.005, **** P < 0.0001. Scale bar: 50 μm ( A and J ) and 25 μm (inset of J ).

Article Snippet: Slides were treated with 3% H 2 O 2 (5 minutes), blocked with 10% goat serum (30 minutes), and incubated with primary antibodies against TCF4 (HPA025958-100UL, Sigma-Aldrich), IL-17C (AF1234, R&D Systems), and rabbit IgG isotype control (NI01, Sigma-Aldrich) overnight at 4°C.

Techniques: Expressing, Staining, Isolation, Control, Enzyme-linked Immunosorbent Assay, Gene Expression

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

Article Title: IL-17C/IL-17RE Augments T Cell Function in Autoimmune Hepatitis.

doi: 10.4049/jimmunol.1600977

Figure Lengend Snippet: FIGURE 3. IL-17C expression is upregulated in autoimmune hepatitis. (A and B) WT mice were intravenously injected with 12 mg/kg Con A. (A) Relative mRNA expression of IL-17C at different time points in liver of Con A–treated WT mice, n = 5 for each group. (B and C) Representative image of liver sections from control and Con A–treated mice (B) or angioma and AIH patients (C) that were stained for IL-17C by immunohistochemistry. Scale bars, 100 mm (left and middle) and 50 mm (right), respectively. (D) Representative images of liver sections from control and Con A–treated mice (upper panel) or angioma and AIH patients (lower panel) that were stained for IL-17RE by immunohistochemistry. Scale bars, 100 mm (left and middle) and 50 mm (right), respectively. (E) IL-17C mRNA expression in different cell types isolated from liver of normal (n = 4) or Con A–treated mice (n = 6) at 8 h post injection. (F) IL-17C mRNA expression in liver of WTand Rag12/2 mice 8 h after Con A injection, n = 4–5 mice per group. (G) IL-17C mRNA expression in mouse primary hepatocytes treated for 4 h with IL-6 (20 ng/ml), IL-4 (20 ng/ml), IL-1a (10 ng/ml), IL-17A (50 ng/ml), IFN-g (20 ng/ml), IL-1b (10 ng/ml), TNF-a (10 ng/ml). Data shown are representative of two independent experiments. *p , 0.05, **p , 0.01, ***p , 0.001.

Article Snippet: Immunohistochemical staining was performed using a rabbit antimouse IL-17C polyclonal Ab (Bioss) or a goat anti-human IL-17C polyclonal Ab (R&D), rabbit anti-mouse/human IL-17RE polyclonal Ab (Biorbyt), and then incubated with a HRP-labeled secondary Ab and followed by 3,39-diaminobenzidine staining.

Techniques: Expressing, Injection, Control, Staining, Immunohistochemistry, Isolation

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

Article Title: IL-17C/IL-17RE Augments T Cell Function in Autoimmune Hepatitis.

doi: 10.4049/jimmunol.1600977

Figure Lengend Snippet: FIGURE 5. IL-17C is required for T and NK cell activation in Con A–induced hepatitis. WT and IL-17C2/2 mice were injected with 12 mg/kg Con A intravenously, and liver MNCs were isolated 24 h later. (A–F) Activation of liver CD4+

Article Snippet: Immunohistochemical staining was performed using a rabbit antimouse IL-17C polyclonal Ab (Bioss) or a goat anti-human IL-17C polyclonal Ab (R&D), rabbit anti-mouse/human IL-17RE polyclonal Ab (Biorbyt), and then incubated with a HRP-labeled secondary Ab and followed by 3,39-diaminobenzidine staining.

Techniques: Activation Assay, Injection, Isolation

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

Article Title: IL-17C/IL-17RE Augments T Cell Function in Autoimmune Hepatitis.

doi: 10.4049/jimmunol.1600977

Figure Lengend Snippet: FIGURE 7. IL-17C–dependent liver damage re- quires NK cells. (A) Representative FACS of liver NK cells from WT (n = 7) and IL-17C2/2 (n = 9) mice 24 h after Con A treatment. (B) Real-time RT- PCR analysis of TRAIL mRNA level in sorted liver CD4+T, CD8+T, NK, NKT cells of WT and IL-17C2/2

Article Snippet: Immunohistochemical staining was performed using a rabbit antimouse IL-17C polyclonal Ab (Bioss) or a goat anti-human IL-17C polyclonal Ab (R&D), rabbit anti-mouse/human IL-17RE polyclonal Ab (Biorbyt), and then incubated with a HRP-labeled secondary Ab and followed by 3,39-diaminobenzidine staining.

Techniques: Quantitative RT-PCR