Fig 1: Effect of IL-22R1 knockdown or overexpression on the proliferation and collagen synthesis of MRC-5 cells. (A) Expression of IL-22R1, COL1a1 and COL1a2 proteins in MRC-5 cells transfected with siR-IL-22R1, siR-NC, an IL-22R1 overexpression plasmid or an NC plasmid, incubated with serum from children with asthma (asthma serum). Western blotting was performed to determine the levels of protein expression. *P<0.05 vs. siR-NC; #P<0.05 vs. NC. (B) Proliferation of MRC-5 cells transfected with siR-IL-22R1, siR-NC, an IL-22R1 overexpression plasmid or an NC plasmid, incubated with asthma serum. The Cell Counting Kit-8 assay was performed to study cell proliferation. *P<0.05 vs. siR-NC; #P<0.05 vs. NC. (C) Cell cycle distribution analysis of MRC-5 cells transfected with siR-IL-22R1, siR-NC, an IL-22R1 overexpression plasmid or an NC plasmid, incubated with asthma serum. Flow cytometry was performed to assess the cell cycle distribution of MRC-5 cells. *P<0.05 vs. siR-NC of the same cell cycle phase; #P<0.05 vs. NC of the same cell cycle phase. IL, interleukin; IL-22R1, IL-22 receptor 1; COL1a1, collagen type I a1 chain; COL1a2, collagen type I a2 chain; siR, small-interfering RNA; NC, negative control.
Fig 2: IL-22 antibody rescued the regulatory effect of serum derived from children with asthma on the biological functions of MRC-5 cells. Cells in the control group were not treated with serum or IL-22 antibody. (A) Proliferation of MRC-5 cells incubated with serum from children with asthma (asthma serum), in the presence or absence of IL-22 antibody. The Cell Counting Kit-8 assay was performed to study cell proliferation. *P<0.05 vs. control. (B) Cell cycle distribution analysis of MRC-5 cells incubated with asthma serum, in the presence or absence of IL-22 antibody. Flow cytometry was performed to assess the cell cycle distribution of MRC-5 cells. *P<0.05 vs. asthma serum group of the same cell cycle phase. (C) Expression of IL-22R1, COL1a1 and COL1a2 proteins in MRC-5 cells incubated with asthma serum, in the presence or absence of IL-22 antibody. Western blotting was performed to determine the levels of protein expression. *P<0.05 vs. control of the same protein; #P<0.05 vs. asthma serum group of the same protein. IL, interleukin; ab, antibody; IL-22R1, IL-22 receptor 1; COL1a1, collagen type I a1 chain; COL1a2, collagen type I a2 chain.
Fig 3: Effect of IL-22+ mononuclear lymphocytes on the biological functions of MRC-5 cells. Cells in the NC group were untreated. (A) Ratio of IL-22+ mononuclear lymphocytes in peripheral blood from healthy children and children with asthma, as determined by flow cytometry. *P<0.05 vs. healthy children. (B) Proliferation of MRC-5 cells co-cultured with mononuclear lymphocytes derived from children with asthma, in the presence or absence of IL-22 antibody. The Cell Counting Kit-8 assay was performed to study cell proliferation. *P<0.05 vs. NC. (C) Cell cycle distribution analysis of MRC-5 cells co-cultured with mononuclear lymphocytes derived from children with asthma, in the presence or absence of IL-22 antibody. Flow cytometry was performed to assess the cell cycle distribution of MRC-5 cells. *P<0.05 vs. NC group of the same cell cycle phase. (D) Expression of COL1a1 and COL1a2 proteins in MRC-5 cells co-cultured with mononuclear lymphocytes derived from children with asthma, in the presence or absence of IL-22 antibody. Western blotting was performed to determine the levels of protein expression. *P<0.05 vs. NC of the same protein. IL, interleukin; NC, negative control; COL1a1, collagen type I a1 chain; COL1a2, collagen type I a2 chain; SSC, side scatter; FSC forward scatter.
Fig 4: Upregulated IL-22 expression in asthma promotes the proliferation and collagen synthesis of MRC-5 cells. (A) Content of IL-22 in serum from healthy children (healthy serum) and children with asthma (asthma serum), determined by ELISA. *P<0.05, as indicated. (B) Expression of IL-22 mRNA in untreated MRC-5 cells and MRC-5 cells treated for 24 h with healthy or asthma serum. Reverse transcription-quantitative PCR was performed to determine IL-22 mRNA expression. *P<0.05, as indicated. (C) Proliferation of MRC-5 cells incubated with healthy or asthma serum. The Cell Counting Kit-8 assay was peformed to study cell proliferation. *P<0.05 vs. control. (D) Cell cycle distribution analysis of MRC-5 cells incubated with healthy or asthma serum. Flow cytometry was performed to assess the cell cycle distribution of MRC-5 cells. *P<0.05 vs. control of the same cell cycle phase. (E) Expression of IL-22R1, COL1a1 and COL1a2 proteins in MRC-5 cells incubated with healthy or asthma serum. Western blotting was performed to determine the levels of protein expression. *P<0.05 vs. control of the same protein. IL, interleukin; IL-22R1, IL-22 receptor 1; COL1a1, collagen type I a1 chain; COL1a2, collagen type I a2 chain.
Fig 5: IL-22/IL-22R1 interaction regulates the biological functions of MRC-5 cells via the JAK/STAT3 signaling pathway. Cells in the NC group were untreated. (A) Proliferation of MRC-5 cells overexpressing IL-22R1, in the presence or absence of the JAK/STAT3 signaling pathway inhibitor stattic (2 µM). The Cell Counting Kit-8 assay was performed to study cell proliferation. *P<0.05 vs. NC. (B) Expression of COL1a1 and COL1a2 proteins in MRC-5 cells overexpressing IL-22R1 in the presence or absence of stattic (2 µM). Western blotting was performed to determine the levels of protein expression. *P<0.05 vs. NC of the same protein. (C) Cell cycle distribution analysis of MRC-5 cells overexpressing IL-22R1 in the presence or absence of stattic (2 µM). Flow cytometry was performed to assess the cell cycle distribution of MRC-5 cells. *P<0.05 vs. NC of the same cell cycle phase. IL, interleukin; IL-22R1, IL-22 receptor 1; NC, negative control; COL1a1, collagen type I a1 chain; COL1a2, collagen type I a2 chain.
Supplier Page from Abcam for Human IL-22 ELISA Kit