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. 2019 Feb 8;13:581–588. doi: 10.2147/DDDT.S194765

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© 2019 Guo et al. This work is published and licensed by Dove Medical Press Limited

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The mRNA and protein detection of CXCL-1 and CXCL-8 using real-time quantitative PCR and ELISA; detection of cleavage effect of CG on FL-IL-36γ using Western blotting.

Notes: (A) HaCaT cells treated with IL-36γ combined with different doses of NE or CG for 24 hours show 100 ng/mL CG used with FL-IL-36γ had synergistic effect on CXCL-1 and CXCL-8 mRNA expression in HaCaT cells. T-IL-36 at 100 ng/mL was used as positive control for IL-36 activity. (B) ELISA analysis of the supernatant confirms CXCL-1 and CXCL-8 expression at the protein level. (C) Western blotting shows that purified CG can cleave FL-IL-36γ, size from 18.7 to 17 KDa. The normalized data are from representative experiment conducted in triplicate. Statistical significance indicated: *P<0.05, ***P<0.001.

Abbreviations: CG, cathepsin G; FL-IL-36γ, full-length-IL-36γ; NE, neutrophil elastase; T-IL-36, truncated IL-36γ.