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Knocking down ADAM‐17 inhibits expression of IL‐6 and TNF‐α. (A and B) Three lentivirus shRNAs (sh‐ADAM17#1, sh‐ADAM17#2, or sh‐ADAM17#3) were introduced into RAW264.7 cells, and the mRNA and protein expression levels of ADAM‐17 measured by WB and qRT‐PCR. (C) NC and sh‐ADAM17#1 RAW264.7 cells were treated with LCWE (1 μg/mL) for 12 h, the protein expression levels of IL‐6, IL‐1β, MCP‐1, and TNF‐α were assayed by ELISA. (D) The mRNA expression level of Areg was measured by qRT‐PCR. (E–H) LCWE‐stimulated RAW264.7 cells were treated with or without exogenous rm‐Areg (1 μg/mL), anti‐Areg (5 μg/mL), or IgG (5 μg/mL) for 12 h, then the protein expression levels of IL‐6, IL‐1β, MCP‐1, and TNF‐α were assayed by ELISA. Results are expressed as the mean ± SD, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Journal: Immunity, Inflammation and Disease

Article Title: Amphiregulin Promotes Proliferation and Migration of the Damaged Endothelial Cells in Kawasaki Disease Cell Models

doi: 10.1002/iid3.70223

Figure Lengend Snippet: Knocking down ADAM‐17 inhibits expression of IL‐6 and TNF‐α. (A and B) Three lentivirus shRNAs (sh‐ADAM17#1, sh‐ADAM17#2, or sh‐ADAM17#3) were introduced into RAW264.7 cells, and the mRNA and protein expression levels of ADAM‐17 measured by WB and qRT‐PCR. (C) NC and sh‐ADAM17#1 RAW264.7 cells were treated with LCWE (1 μg/mL) for 12 h, the protein expression levels of IL‐6, IL‐1β, MCP‐1, and TNF‐α were assayed by ELISA. (D) The mRNA expression level of Areg was measured by qRT‐PCR. (E–H) LCWE‐stimulated RAW264.7 cells were treated with or without exogenous rm‐Areg (1 μg/mL), anti‐Areg (5 μg/mL), or IgG (5 μg/mL) for 12 h, then the protein expression levels of IL‐6, IL‐1β, MCP‐1, and TNF‐α were assayed by ELISA. Results are expressed as the mean ± SD, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Primary antibodies against GAPDH (Cat. No. 60004‐1‐Ig) and ADAM‐17 (Cat. No. 29948‐1‐AP) were purchased from Proteintech (Wuhan, China).

Techniques: Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay