il−18 Search Results


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Figure 3. Silencing p62 reduces IL‑18 secretion and promotes cell survival of macrophages with prolonged oxLDL exposure. (A) THP‑M cells treated with oxLDL (20 µg/ml) for the indicated time following transfection with 50 nM con siRNA or p62 siRNA. Western blotting experiments were performed on total protein extracts using anti‑p62 antibody, and β‑actin expression was used as a loading control. The graph represents the values of p62 band intensity after normalization to β‑actin by densitometry. *P<0.05 vs. 24 h group. (B) The graph represents values of the SQSTM1 mRNA expression following normalization for β‑actin mRNA by reverse transcription‑quantitative polymerase chain reaction. *P<0.05 vs. 24 h group. (C) <t>ELISA</t> of secreted IL‑18 in the media of cells treated with con or p62 siRNA and 20 µg/ml oxLDL for 0, 12, 24, 48 and 72 h. *P<0.05 and **P<0.01 vs. 0 h; ##P<0.01 as indicated. (D) THP‑M cells were pretreated with con or p62 siRNA and then treated with 20 µg/ml oxLDL for the indicated times. Cell viability was analyzed by the MTT assay. Results are expressed as a percentage of con siRNA group at 0 h. *P<0.05 vs. con siRNA group. (E) Determination of cell death by flow cytometry of Annexin V‑fluorescein isothiocyanate/PI staining in THP‑M cells transfected with con or p62 siRNA and then incubated with oxLDL (20 µg/ml) for the indicated times. Data are representatives of three independent experiments. **P<0.01 as indicated. <t>IL,</t> <t>interleukin;</t> oxLDL, oxidized low‑density lipoprotein; THP‑M, THP‑1‑derived macrophages; siRNA, small interfering RNA; con, control; PI, propidium iodide.
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Figure 3. Silencing p62 reduces IL‑18 secretion and promotes cell survival of macrophages with prolonged oxLDL exposure. (A) THP‑M cells treated with oxLDL (20 µg/ml) for the indicated time following transfection with 50 nM con siRNA or p62 siRNA. Western blotting experiments were performed on total protein extracts using anti‑p62 antibody, and β‑actin expression was used as a loading control. The graph represents the values of p62 band intensity after normalization to β‑actin by densitometry. *P<0.05 vs. 24 h group. (B) The graph represents values of the SQSTM1 mRNA expression following normalization for β‑actin mRNA by reverse transcription‑quantitative polymerase chain reaction. *P<0.05 vs. 24 h group. (C) <t>ELISA</t> of secreted IL‑18 in the media of cells treated with con or p62 siRNA and 20 µg/ml oxLDL for 0, 12, 24, 48 and 72 h. *P<0.05 and **P<0.01 vs. 0 h; ##P<0.01 as indicated. (D) THP‑M cells were pretreated with con or p62 siRNA and then treated with 20 µg/ml oxLDL for the indicated times. Cell viability was analyzed by the MTT assay. Results are expressed as a percentage of con siRNA group at 0 h. *P<0.05 vs. con siRNA group. (E) Determination of cell death by flow cytometry of Annexin V‑fluorescein isothiocyanate/PI staining in THP‑M cells transfected with con or p62 siRNA and then incubated with oxLDL (20 µg/ml) for the indicated times. Data are representatives of three independent experiments. **P<0.01 as indicated. <t>IL,</t> <t>interleukin;</t> oxLDL, oxidized low‑density lipoprotein; THP‑M, THP‑1‑derived macrophages; siRNA, small interfering RNA; con, control; PI, propidium iodide.
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Figure 3. Silencing p62 reduces IL‑18 secretion and promotes cell survival of macrophages with prolonged oxLDL exposure. (A) THP‑M cells treated with oxLDL (20 µg/ml) for the indicated time following transfection with 50 nM con siRNA or p62 siRNA. Western blotting experiments were performed on total protein extracts using anti‑p62 antibody, and β‑actin expression was used as a loading control. The graph represents the values of p62 band intensity after normalization to β‑actin by densitometry. *P<0.05 vs. 24 h group. (B) The graph represents values of the SQSTM1 mRNA expression following normalization for β‑actin mRNA by reverse transcription‑quantitative polymerase chain reaction. *P<0.05 vs. 24 h group. (C) <t>ELISA</t> of secreted IL‑18 in the media of cells treated with con or p62 siRNA and 20 µg/ml oxLDL for 0, 12, 24, 48 and 72 h. *P<0.05 and **P<0.01 vs. 0 h; ##P<0.01 as indicated. (D) THP‑M cells were pretreated with con or p62 siRNA and then treated with 20 µg/ml oxLDL for the indicated times. Cell viability was analyzed by the MTT assay. Results are expressed as a percentage of con siRNA group at 0 h. *P<0.05 vs. con siRNA group. (E) Determination of cell death by flow cytometry of Annexin V‑fluorescein isothiocyanate/PI staining in THP‑M cells transfected with con or p62 siRNA and then incubated with oxLDL (20 µg/ml) for the indicated times. Data are representatives of three independent experiments. **P<0.01 as indicated. <t>IL,</t> <t>interleukin;</t> oxLDL, oxidized low‑density lipoprotein; THP‑M, THP‑1‑derived macrophages; siRNA, small interfering RNA; con, control; PI, propidium iodide.
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Figure 3. Silencing p62 reduces IL‑18 secretion and promotes cell survival of macrophages with prolonged oxLDL exposure. (A) THP‑M cells treated with oxLDL (20 µg/ml) for the indicated time following transfection with 50 nM con siRNA or p62 siRNA. Western blotting experiments were performed on total protein extracts using anti‑p62 antibody, and β‑actin expression was used as a loading control. The graph represents the values of p62 band intensity after normalization to β‑actin by densitometry. *P<0.05 vs. 24 h group. (B) The graph represents values of the SQSTM1 mRNA expression following normalization for β‑actin mRNA by reverse transcription‑quantitative polymerase chain reaction. *P<0.05 vs. 24 h group. (C) <t>ELISA</t> of secreted IL‑18 in the media of cells treated with con or p62 siRNA and 20 µg/ml oxLDL for 0, 12, 24, 48 and 72 h. *P<0.05 and **P<0.01 vs. 0 h; ##P<0.01 as indicated. (D) THP‑M cells were pretreated with con or p62 siRNA and then treated with 20 µg/ml oxLDL for the indicated times. Cell viability was analyzed by the MTT assay. Results are expressed as a percentage of con siRNA group at 0 h. *P<0.05 vs. con siRNA group. (E) Determination of cell death by flow cytometry of Annexin V‑fluorescein isothiocyanate/PI staining in THP‑M cells transfected with con or p62 siRNA and then incubated with oxLDL (20 µg/ml) for the indicated times. Data are representatives of three independent experiments. **P<0.01 as indicated. <t>IL,</t> <t>interleukin;</t> oxLDL, oxidized low‑density lipoprotein; THP‑M, THP‑1‑derived macrophages; siRNA, small interfering RNA; con, control; PI, propidium iodide.
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Figure 3. Silencing p62 reduces IL‑18 secretion and promotes cell survival of macrophages with prolonged oxLDL exposure. (A) THP‑M cells treated with oxLDL (20 µg/ml) for the indicated time following transfection with 50 nM con siRNA or p62 siRNA. Western blotting experiments were performed on total protein extracts using anti‑p62 antibody, and β‑actin expression was used as a loading control. The graph represents the values of p62 band intensity after normalization to β‑actin by densitometry. *P<0.05 vs. 24 h group. (B) The graph represents values of the SQSTM1 mRNA expression following normalization for β‑actin mRNA by reverse transcription‑quantitative polymerase chain reaction. *P<0.05 vs. 24 h group. (C) <t>ELISA</t> of secreted IL‑18 in the media of cells treated with con or p62 siRNA and 20 µg/ml oxLDL for 0, 12, 24, 48 and 72 h. *P<0.05 and **P<0.01 vs. 0 h; ##P<0.01 as indicated. (D) THP‑M cells were pretreated with con or p62 siRNA and then treated with 20 µg/ml oxLDL for the indicated times. Cell viability was analyzed by the MTT assay. Results are expressed as a percentage of con siRNA group at 0 h. *P<0.05 vs. con siRNA group. (E) Determination of cell death by flow cytometry of Annexin V‑fluorescein isothiocyanate/PI staining in THP‑M cells transfected with con or p62 siRNA and then incubated with oxLDL (20 µg/ml) for the indicated times. Data are representatives of three independent experiments. **P<0.01 as indicated. <t>IL,</t> <t>interleukin;</t> oxLDL, oxidized low‑density lipoprotein; THP‑M, THP‑1‑derived macrophages; siRNA, small interfering RNA; con, control; PI, propidium iodide.
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Figure 3. Silencing p62 reduces IL‑18 secretion and promotes cell survival of macrophages with prolonged oxLDL exposure. (A) THP‑M cells treated with oxLDL (20 µg/ml) for the indicated time following transfection with 50 nM con siRNA or p62 siRNA. Western blotting experiments were performed on total protein extracts using anti‑p62 antibody, and β‑actin expression was used as a loading control. The graph represents the values of p62 band intensity after normalization to β‑actin by densitometry. *P<0.05 vs. 24 h group. (B) The graph represents values of the SQSTM1 mRNA expression following normalization for β‑actin mRNA by reverse transcription‑quantitative polymerase chain reaction. *P<0.05 vs. 24 h group. (C) <t>ELISA</t> of secreted IL‑18 in the media of cells treated with con or p62 siRNA and 20 µg/ml oxLDL for 0, 12, 24, 48 and 72 h. *P<0.05 and **P<0.01 vs. 0 h; ##P<0.01 as indicated. (D) THP‑M cells were pretreated with con or p62 siRNA and then treated with 20 µg/ml oxLDL for the indicated times. Cell viability was analyzed by the MTT assay. Results are expressed as a percentage of con siRNA group at 0 h. *P<0.05 vs. con siRNA group. (E) Determination of cell death by flow cytometry of Annexin V‑fluorescein isothiocyanate/PI staining in THP‑M cells transfected with con or p62 siRNA and then incubated with oxLDL (20 µg/ml) for the indicated times. Data are representatives of three independent experiments. **P<0.01 as indicated. <t>IL,</t> <t>interleukin;</t> oxLDL, oxidized low‑density lipoprotein; THP‑M, THP‑1‑derived macrophages; siRNA, small interfering RNA; con, control; PI, propidium iodide.
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Figure 3. Silencing p62 reduces IL‑18 secretion and promotes cell survival of macrophages with prolonged oxLDL exposure. (A) THP‑M cells treated with oxLDL (20 µg/ml) for the indicated time following transfection with 50 nM con siRNA or p62 siRNA. Western blotting experiments were performed on total protein extracts using anti‑p62 antibody, and β‑actin expression was used as a loading control. The graph represents the values of p62 band intensity after normalization to β‑actin by densitometry. *P<0.05 vs. 24 h group. (B) The graph represents values of the SQSTM1 mRNA expression following normalization for β‑actin mRNA by reverse transcription‑quantitative polymerase chain reaction. *P<0.05 vs. 24 h group. (C) <t>ELISA</t> of secreted IL‑18 in the media of cells treated with con or p62 siRNA and 20 µg/ml oxLDL for 0, 12, 24, 48 and 72 h. *P<0.05 and **P<0.01 vs. 0 h; ##P<0.01 as indicated. (D) THP‑M cells were pretreated with con or p62 siRNA and then treated with 20 µg/ml oxLDL for the indicated times. Cell viability was analyzed by the MTT assay. Results are expressed as a percentage of con siRNA group at 0 h. *P<0.05 vs. con siRNA group. (E) Determination of cell death by flow cytometry of Annexin V‑fluorescein isothiocyanate/PI staining in THP‑M cells transfected with con or p62 siRNA and then incubated with oxLDL (20 µg/ml) for the indicated times. Data are representatives of three independent experiments. **P<0.01 as indicated. <t>IL,</t> <t>interleukin;</t> oxLDL, oxidized low‑density lipoprotein; THP‑M, THP‑1‑derived macrophages; siRNA, small interfering RNA; con, control; PI, propidium iodide.
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Figure 3. Silencing p62 reduces IL‑18 secretion and promotes cell survival of macrophages with prolonged oxLDL exposure. (A) THP‑M cells treated with oxLDL (20 µg/ml) for the indicated time following transfection with 50 nM con siRNA or p62 siRNA. Western blotting experiments were performed on total protein extracts using anti‑p62 antibody, and β‑actin expression was used as a loading control. The graph represents the values of p62 band intensity after normalization to β‑actin by densitometry. *P<0.05 vs. 24 h group. (B) The graph represents values of the SQSTM1 mRNA expression following normalization for β‑actin mRNA by reverse transcription‑quantitative polymerase chain reaction. *P<0.05 vs. 24 h group. (C) <t>ELISA</t> of secreted IL‑18 in the media of cells treated with con or p62 siRNA and 20 µg/ml oxLDL for 0, 12, 24, 48 and 72 h. *P<0.05 and **P<0.01 vs. 0 h; ##P<0.01 as indicated. (D) THP‑M cells were pretreated with con or p62 siRNA and then treated with 20 µg/ml oxLDL for the indicated times. Cell viability was analyzed by the MTT assay. Results are expressed as a percentage of con siRNA group at 0 h. *P<0.05 vs. con siRNA group. (E) Determination of cell death by flow cytometry of Annexin V‑fluorescein isothiocyanate/PI staining in THP‑M cells transfected with con or p62 siRNA and then incubated with oxLDL (20 µg/ml) for the indicated times. Data are representatives of three independent experiments. **P<0.01 as indicated. <t>IL,</t> <t>interleukin;</t> oxLDL, oxidized low‑density lipoprotein; THP‑M, THP‑1‑derived macrophages; siRNA, small interfering RNA; con, control; PI, propidium iodide.
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Figure 3. Silencing p62 reduces IL‑18 secretion and promotes cell survival of macrophages with prolonged oxLDL exposure. (A) THP‑M cells treated with oxLDL (20 µg/ml) for the indicated time following transfection with 50 nM con siRNA or p62 siRNA. Western blotting experiments were performed on total protein extracts using anti‑p62 antibody, and β‑actin expression was used as a loading control. The graph represents the values of p62 band intensity after normalization to β‑actin by densitometry. *P<0.05 vs. 24 h group. (B) The graph represents values of the SQSTM1 mRNA expression following normalization for β‑actin mRNA by reverse transcription‑quantitative polymerase chain reaction. *P<0.05 vs. 24 h group. (C) <t>ELISA</t> of secreted IL‑18 in the media of cells treated with con or p62 siRNA and 20 µg/ml oxLDL for 0, 12, 24, 48 and 72 h. *P<0.05 and **P<0.01 vs. 0 h; ##P<0.01 as indicated. (D) THP‑M cells were pretreated with con or p62 siRNA and then treated with 20 µg/ml oxLDL for the indicated times. Cell viability was analyzed by the MTT assay. Results are expressed as a percentage of con siRNA group at 0 h. *P<0.05 vs. con siRNA group. (E) Determination of cell death by flow cytometry of Annexin V‑fluorescein isothiocyanate/PI staining in THP‑M cells transfected with con or p62 siRNA and then incubated with oxLDL (20 µg/ml) for the indicated times. Data are representatives of three independent experiments. **P<0.01 as indicated. <t>IL,</t> <t>interleukin;</t> oxLDL, oxidized low‑density lipoprotein; THP‑M, THP‑1‑derived macrophages; siRNA, small interfering RNA; con, control; PI, propidium iodide.
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Figure 3. Silencing p62 reduces IL‑18 secretion and promotes cell survival of macrophages with prolonged oxLDL exposure. (A) THP‑M cells treated with oxLDL (20 µg/ml) for the indicated time following transfection with 50 nM con siRNA or p62 siRNA. Western blotting experiments were performed on total protein extracts using anti‑p62 antibody, and β‑actin expression was used as a loading control. The graph represents the values of p62 band intensity after normalization to β‑actin by densitometry. *P<0.05 vs. 24 h group. (B) The graph represents values of the SQSTM1 mRNA expression following normalization for β‑actin mRNA by reverse transcription‑quantitative polymerase chain reaction. *P<0.05 vs. 24 h group. (C) <t>ELISA</t> of secreted IL‑18 in the media of cells treated with con or p62 siRNA and 20 µg/ml oxLDL for 0, 12, 24, 48 and 72 h. *P<0.05 and **P<0.01 vs. 0 h; ##P<0.01 as indicated. (D) THP‑M cells were pretreated with con or p62 siRNA and then treated with 20 µg/ml oxLDL for the indicated times. Cell viability was analyzed by the MTT assay. Results are expressed as a percentage of con siRNA group at 0 h. *P<0.05 vs. con siRNA group. (E) Determination of cell death by flow cytometry of Annexin V‑fluorescein isothiocyanate/PI staining in THP‑M cells transfected with con or p62 siRNA and then incubated with oxLDL (20 µg/ml) for the indicated times. Data are representatives of three independent experiments. **P<0.01 as indicated. <t>IL,</t> <t>interleukin;</t> oxLDL, oxidized low‑density lipoprotein; THP‑M, THP‑1‑derived macrophages; siRNA, small interfering RNA; con, control; PI, propidium iodide.
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Figure 3. Silencing p62 reduces IL‑18 secretion and promotes cell survival of macrophages with prolonged oxLDL exposure. (A) THP‑M cells treated with oxLDL (20 µg/ml) for the indicated time following transfection with 50 nM con siRNA or p62 siRNA. Western blotting experiments were performed on total protein extracts using anti‑p62 antibody, and β‑actin expression was used as a loading control. The graph represents the values of p62 band intensity after normalization to β‑actin by densitometry. *P<0.05 vs. 24 h group. (B) The graph represents values of the SQSTM1 mRNA expression following normalization for β‑actin mRNA by reverse transcription‑quantitative polymerase chain reaction. *P<0.05 vs. 24 h group. (C) <t>ELISA</t> of secreted IL‑18 in the media of cells treated with con or p62 siRNA and 20 µg/ml oxLDL for 0, 12, 24, 48 and 72 h. *P<0.05 and **P<0.01 vs. 0 h; ##P<0.01 as indicated. (D) THP‑M cells were pretreated with con or p62 siRNA and then treated with 20 µg/ml oxLDL for the indicated times. Cell viability was analyzed by the MTT assay. Results are expressed as a percentage of con siRNA group at 0 h. *P<0.05 vs. con siRNA group. (E) Determination of cell death by flow cytometry of Annexin V‑fluorescein isothiocyanate/PI staining in THP‑M cells transfected with con or p62 siRNA and then incubated with oxLDL (20 µg/ml) for the indicated times. Data are representatives of three independent experiments. **P<0.01 as indicated. <t>IL,</t> <t>interleukin;</t> oxLDL, oxidized low‑density lipoprotein; THP‑M, THP‑1‑derived macrophages; siRNA, small interfering RNA; con, control; PI, propidium iodide.
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Figure 3. Silencing p62 reduces IL‑18 secretion and promotes cell survival of macrophages with prolonged oxLDL exposure. (A) THP‑M cells treated with oxLDL (20 µg/ml) for the indicated time following transfection with 50 nM con siRNA or p62 siRNA. Western blotting experiments were performed on total protein extracts using anti‑p62 antibody, and β‑actin expression was used as a loading control. The graph represents the values of p62 band intensity after normalization to β‑actin by densitometry. *P<0.05 vs. 24 h group. (B) The graph represents values of the SQSTM1 mRNA expression following normalization for β‑actin mRNA by reverse transcription‑quantitative polymerase chain reaction. *P<0.05 vs. 24 h group. (C) <t>ELISA</t> of secreted IL‑18 in the media of cells treated with con or p62 siRNA and 20 µg/ml oxLDL for 0, 12, 24, 48 and 72 h. *P<0.05 and **P<0.01 vs. 0 h; ##P<0.01 as indicated. (D) THP‑M cells were pretreated with con or p62 siRNA and then treated with 20 µg/ml oxLDL for the indicated times. Cell viability was analyzed by the MTT assay. Results are expressed as a percentage of con siRNA group at 0 h. *P<0.05 vs. con siRNA group. (E) Determination of cell death by flow cytometry of Annexin V‑fluorescein isothiocyanate/PI staining in THP‑M cells transfected with con or p62 siRNA and then incubated with oxLDL (20 µg/ml) for the indicated times. Data are representatives of three independent experiments. **P<0.01 as indicated. <t>IL,</t> <t>interleukin;</t> oxLDL, oxidized low‑density lipoprotein; THP‑M, THP‑1‑derived macrophages; siRNA, small interfering RNA; con, control; PI, propidium iodide.
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Image Search Results


Figure 3. Silencing p62 reduces IL‑18 secretion and promotes cell survival of macrophages with prolonged oxLDL exposure. (A) THP‑M cells treated with oxLDL (20 µg/ml) for the indicated time following transfection with 50 nM con siRNA or p62 siRNA. Western blotting experiments were performed on total protein extracts using anti‑p62 antibody, and β‑actin expression was used as a loading control. The graph represents the values of p62 band intensity after normalization to β‑actin by densitometry. *P<0.05 vs. 24 h group. (B) The graph represents values of the SQSTM1 mRNA expression following normalization for β‑actin mRNA by reverse transcription‑quantitative polymerase chain reaction. *P<0.05 vs. 24 h group. (C) ELISA of secreted IL‑18 in the media of cells treated with con or p62 siRNA and 20 µg/ml oxLDL for 0, 12, 24, 48 and 72 h. *P<0.05 and **P<0.01 vs. 0 h; ##P<0.01 as indicated. (D) THP‑M cells were pretreated with con or p62 siRNA and then treated with 20 µg/ml oxLDL for the indicated times. Cell viability was analyzed by the MTT assay. Results are expressed as a percentage of con siRNA group at 0 h. *P<0.05 vs. con siRNA group. (E) Determination of cell death by flow cytometry of Annexin V‑fluorescein isothiocyanate/PI staining in THP‑M cells transfected with con or p62 siRNA and then incubated with oxLDL (20 µg/ml) for the indicated times. Data are representatives of three independent experiments. **P<0.01 as indicated. IL, interleukin; oxLDL, oxidized low‑density lipoprotein; THP‑M, THP‑1‑derived macrophages; siRNA, small interfering RNA; con, control; PI, propidium iodide.

Journal: Experimental and therapeutic medicine

Article Title: Oxidized low-density lipoprotein-induced p62/SQSTM1 accumulation in THP-1-derived macrophages promotes IL-18 secretion and cell death.

doi: 10.3892/etm.2017.5221

Figure Lengend Snippet: Figure 3. Silencing p62 reduces IL‑18 secretion and promotes cell survival of macrophages with prolonged oxLDL exposure. (A) THP‑M cells treated with oxLDL (20 µg/ml) for the indicated time following transfection with 50 nM con siRNA or p62 siRNA. Western blotting experiments were performed on total protein extracts using anti‑p62 antibody, and β‑actin expression was used as a loading control. The graph represents the values of p62 band intensity after normalization to β‑actin by densitometry. *P<0.05 vs. 24 h group. (B) The graph represents values of the SQSTM1 mRNA expression following normalization for β‑actin mRNA by reverse transcription‑quantitative polymerase chain reaction. *P<0.05 vs. 24 h group. (C) ELISA of secreted IL‑18 in the media of cells treated with con or p62 siRNA and 20 µg/ml oxLDL for 0, 12, 24, 48 and 72 h. *P<0.05 and **P<0.01 vs. 0 h; ##P<0.01 as indicated. (D) THP‑M cells were pretreated with con or p62 siRNA and then treated with 20 µg/ml oxLDL for the indicated times. Cell viability was analyzed by the MTT assay. Results are expressed as a percentage of con siRNA group at 0 h. *P<0.05 vs. con siRNA group. (E) Determination of cell death by flow cytometry of Annexin V‑fluorescein isothiocyanate/PI staining in THP‑M cells transfected with con or p62 siRNA and then incubated with oxLDL (20 µg/ml) for the indicated times. Data are representatives of three independent experiments. **P<0.01 as indicated. IL, interleukin; oxLDL, oxidized low‑density lipoprotein; THP‑M, THP‑1‑derived macrophages; siRNA, small interfering RNA; con, control; PI, propidium iodide.

Article Snippet: Interleukin-18 (IL-18) levels from culture media was measured with an ELISA kit from R&D Systems, Inc., (cat. no. 7620; Minneapolis, MN, USA), according to the manufacturer's instructions.

Techniques: Transfection, Western Blot, Expressing, Control, Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, MTT Assay, Flow Cytometry, Staining, Incubation, Small Interfering RNA