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Image Search Results
Journal: Journal of Biological Chemistry
Article Title: Dynamic Modulation of Innate Immune Response by Varying Dosages of Lipopolysaccharide (LPS) in Human Monocytic Cells
doi: 10.1074/jbc.m114.583518
Figure Lengend Snippet: FIGURE 4. Differential regulation of FoxO1 and CREB by LPS. THP-1 cells weretreatedwitheither50pg/mlor100ng/mlLPSforvarioustimeperiodsas indicated. Whole cell lysates were separated on SDS-PAGE. The levels of pFoxO1 and total FoxO1 (A) and pCREB-S133 and total CREB (B) were deter- mined by Western blot with specific antibodies. Data are representative of three separate experiments.
Article Snippet: Protein samples were run on 10% acrylamide gels at 100 V, followed by transfer at 110 V before blocking in 5% milk in TBS-T. Antibodies against pCREB-S133 (Cell Signaling 9191) were used at a concentration of 1:4000 in 5% milk in TBS-T, and CREB (Cell Signaling 9197S),
Techniques: SDS Page, Western Blot
Journal: Journal of Biological Chemistry
Article Title: Dynamic Modulation of Innate Immune Response by Varying Dosages of Lipopolysaccharide (LPS) in Human Monocytic Cells
doi: 10.1074/jbc.m114.583518
Figure Lengend Snippet: FIGURE 4. Differential regulation of FoxO1 and CREB by LPS. THP-1 cells weretreatedwitheither50pg/mlor100ng/mlLPSforvarioustimeperiodsas indicated. Whole cell lysates were separated on SDS-PAGE. The levels of pFoxO1 and total FoxO1 (A) and pCREB-S133 and total CREB (B) were deter- mined by Western blot with specific antibodies. Data are representative of three separate experiments.
Article Snippet: Protein samples were run on 10% acrylamide gels at 100 V, followed by transfer at 110 V before blocking in 5% milk in TBS-T. Antibodies against pCREB-S133 (Cell Signaling 9191) were used at a concentration of 1:4000 in 5% milk in TBS-T, and CREB (Cell Signaling 9197S), pFoxO1-S256 (Cell Signaling 9461S),
Techniques: SDS Page, Western Blot
Journal: Journal of Biological Chemistry
Article Title: Dynamic Modulation of Innate Immune Response by Varying Dosages of Lipopolysaccharide (LPS) in Human Monocytic Cells
doi: 10.1074/jbc.m114.583518
Figure Lengend Snippet: FIGURE 5. Effect of GSK3 inhibition on cellular responses to LPS stimulation. A, THP-1 cells were treated with the combination of 50 pg/ml LPS alone or togetherwith10Mindirubinfor4h.TotalRNAwasharvestedandusedforreal-timeRT-PCRanalysisforIL-6.B,totalproteinlysateswereharvestedfromTHP-1 cells treated as indicated and used for Western blot analyses for FoxO1 and GAPDH. C, THP-1 cells were treated with the combination of 50 pg/ml LPS alone or together with 10 M SB216763 for 4 h. Total RNA was harvested and used for real-time RT-PCR analysis for IL-33. D, total protein lysates were harvested from THP-1 cells treated as indicated, and used for Western blot analyses for CREB and GAPDH. Data are representative of three separate experiments (*, p 0.05, Student’s t test).
Article Snippet: Protein samples were run on 10% acrylamide gels at 100 V, followed by transfer at 110 V before blocking in 5% milk in TBS-T. Antibodies against pCREB-S133 (Cell Signaling 9191) were used at a concentration of 1:4000 in 5% milk in TBS-T, and CREB (Cell Signaling 9197S), pFoxO1-S256 (Cell Signaling 9461S),
Techniques: Inhibition, Western Blot, Quantitative RT-PCR
Journal: Journal of Biological Chemistry
Article Title: Dynamic Modulation of Innate Immune Response by Varying Dosages of Lipopolysaccharide (LPS) in Human Monocytic Cells
doi: 10.1074/jbc.m114.583518
Figure Lengend Snippet: FIGURE 4. Differential regulation of FoxO1 and CREB by LPS. THP-1 cells weretreatedwitheither50pg/mlor100ng/mlLPSforvarioustimeperiodsas indicated. Whole cell lysates were separated on SDS-PAGE. The levels of pFoxO1 and total FoxO1 (A) and pCREB-S133 and total CREB (B) were deter- mined by Western blot with specific antibodies. Data are representative of three separate experiments.
Article Snippet: Protein samples were run on 10% acrylamide gels at 100 V, followed by transfer at 110 V before blocking in 5% milk in TBS-T. Antibodies against pCREB-S133 (Cell Signaling 9191) were used at a concentration of 1:4000 in 5% milk in TBS-T, and
Techniques: SDS Page, Western Blot
Journal: Journal of Biological Chemistry
Article Title: Dynamic Modulation of Innate Immune Response by Varying Dosages of Lipopolysaccharide (LPS) in Human Monocytic Cells
doi: 10.1074/jbc.m114.583518
Figure Lengend Snippet: FIGURE 5. Effect of GSK3 inhibition on cellular responses to LPS stimulation. A, THP-1 cells were treated with the combination of 50 pg/ml LPS alone or togetherwith10Mindirubinfor4h.TotalRNAwasharvestedandusedforreal-timeRT-PCRanalysisforIL-6.B,totalproteinlysateswereharvestedfromTHP-1 cells treated as indicated and used for Western blot analyses for FoxO1 and GAPDH. C, THP-1 cells were treated with the combination of 50 pg/ml LPS alone or together with 10 M SB216763 for 4 h. Total RNA was harvested and used for real-time RT-PCR analysis for IL-33. D, total protein lysates were harvested from THP-1 cells treated as indicated, and used for Western blot analyses for CREB and GAPDH. Data are representative of three separate experiments (*, p 0.05, Student’s t test).
Article Snippet: Protein samples were run on 10% acrylamide gels at 100 V, followed by transfer at 110 V before blocking in 5% milk in TBS-T. Antibodies against pCREB-S133 (Cell Signaling 9191) were used at a concentration of 1:4000 in 5% milk in TBS-T, and
Techniques: Inhibition, Western Blot, Quantitative RT-PCR
Journal: Journal of Biological Chemistry
Article Title: Dynamic Modulation of Innate Immune Response by Varying Dosages of Lipopolysaccharide (LPS) in Human Monocytic Cells
doi: 10.1074/jbc.m114.583518
Figure Lengend Snippet: FIGURE 3. Differential regulation of GSK3 and Akt in THP-1 cells by LPS. THP-1 cells were treated with either 50 pg/ml or 100 ng/ml LPS for various time periods as indicated. Whole cell lysates were separated on SDS-PAGE. The levels of pAkt-S473 and total Akt (A) pGSK3-Y216 and total GSK3 (B), and pPyk2-Y402 and total Pyk2 (C) were determined by Western blot with specific antibodies. Data are representative of three separate experiments.
Article Snippet: Protein samples were run on 10% acrylamide gels at 100 V, followed by transfer at 110 V before blocking in 5% milk in TBS-T. Antibodies against pCREB-S133 (Cell Signaling 9191) were used at a concentration of 1:4000 in 5% milk in TBS-T, and CREB (Cell Signaling 9197S), pFoxO1-S256 (Cell Signaling 9461S), FoxO1 (Cell Signaling 9454S), Akt (Santa Cruz Biotechnology sc-8312), GSK3 (Santa Cruz Biotechnol- AUGUST 1, 2014 • VOLUME 289 • NUMBER 31 JOURNAL OF BIOLOGICAL CHEMISTRY 21585 ogy sc-9166), and GAPDH (Santa Cruz Biotechnology sc-25778) at 1:1000.
Techniques: SDS Page, Western Blot