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The <t>Akt/mTOR/FOXO1</t> signaling pathway was associated with the antigliomas of XHP. (a) After being incubated with different doses of XHP, 2 µ M PTX or control medium for 24h, U-87 MG cells were lysed and subjected to western blot analysis of Akt, p-Akt, mTOR, p-mTOR, FOXO1, and p-FOXO1 antibodies. (b) Western blot analysis of AKT, p-Akt, mTOR, p-mTOR, FOXO1, and p-FOXO1 expression in U-87 MG cells after treatment with LY294002 (20 μ M), <t>Rapamycin</t> (50 nM), or NAC (5 mM) with or without XHP. (c) Confocal images of U-87 MG cells treated with XHP stained with anti-FOXO1 antibody. (d) Confocal images of U-87 MG cells treated with XHP were stained with anti-p-FOXO1 antibody. (e) Quantitative RT-PCR analysis of U-87 MG cells subjected to FOXO1-specific knockdown, results were normalized with GAPDH level and expressed as folds of control (n = 3, mean ± SD). (f) U-87 MG cells subjected to FOXO1-specific knockdown were assayed by western blot. (g) U-87 MG cells treated with XHP in the absence or presence of LY294002 (20 μ M), Rapamycin (50 nM), or FOXO1-specific siRNA (10 nM). The proportion of Annexin V positive cells was measured by flow cytometry. Data are mean ± SD of three different experiments. ∗P < 0.05, ∗∗P < 0.01 significantly different from the control.
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The <t>Akt/mTOR/FOXO1</t> signaling pathway was associated with the antigliomas of XHP. (a) After being incubated with different doses of XHP, 2 µ M PTX or control medium for 24h, U-87 MG cells were lysed and subjected to western blot analysis of Akt, p-Akt, mTOR, p-mTOR, FOXO1, and p-FOXO1 antibodies. (b) Western blot analysis of AKT, p-Akt, mTOR, p-mTOR, FOXO1, and p-FOXO1 expression in U-87 MG cells after treatment with LY294002 (20 μ M), <t>Rapamycin</t> (50 nM), or NAC (5 mM) with or without XHP. (c) Confocal images of U-87 MG cells treated with XHP stained with anti-FOXO1 antibody. (d) Confocal images of U-87 MG cells treated with XHP were stained with anti-p-FOXO1 antibody. (e) Quantitative RT-PCR analysis of U-87 MG cells subjected to FOXO1-specific knockdown, results were normalized with GAPDH level and expressed as folds of control (n = 3, mean ± SD). (f) U-87 MG cells subjected to FOXO1-specific knockdown were assayed by western blot. (g) U-87 MG cells treated with XHP in the absence or presence of LY294002 (20 μ M), Rapamycin (50 nM), or FOXO1-specific siRNA (10 nM). The proportion of Annexin V positive cells was measured by flow cytometry. Data are mean ± SD of three different experiments. ∗P < 0.05, ∗∗P < 0.01 significantly different from the control.
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The <t>Akt/mTOR/FOXO1</t> signaling pathway was associated with the antigliomas of XHP. (a) After being incubated with different doses of XHP, 2 µ M PTX or control medium for 24h, U-87 MG cells were lysed and subjected to western blot analysis of Akt, p-Akt, mTOR, p-mTOR, FOXO1, and p-FOXO1 antibodies. (b) Western blot analysis of AKT, p-Akt, mTOR, p-mTOR, FOXO1, and p-FOXO1 expression in U-87 MG cells after treatment with LY294002 (20 μ M), <t>Rapamycin</t> (50 nM), or NAC (5 mM) with or without XHP. (c) Confocal images of U-87 MG cells treated with XHP stained with anti-FOXO1 antibody. (d) Confocal images of U-87 MG cells treated with XHP were stained with anti-p-FOXO1 antibody. (e) Quantitative RT-PCR analysis of U-87 MG cells subjected to FOXO1-specific knockdown, results were normalized with GAPDH level and expressed as folds of control (n = 3, mean ± SD). (f) U-87 MG cells subjected to FOXO1-specific knockdown were assayed by western blot. (g) U-87 MG cells treated with XHP in the absence or presence of LY294002 (20 μ M), Rapamycin (50 nM), or FOXO1-specific siRNA (10 nM). The proportion of Annexin V positive cells was measured by flow cytometry. Data are mean ± SD of three different experiments. ∗P < 0.05, ∗∗P < 0.01 significantly different from the control.
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The <t>Akt/mTOR/FOXO1</t> signaling pathway was associated with the antigliomas of XHP. (a) After being incubated with different doses of XHP, 2 µ M PTX or control medium for 24h, U-87 MG cells were lysed and subjected to western blot analysis of Akt, p-Akt, mTOR, p-mTOR, FOXO1, and p-FOXO1 antibodies. (b) Western blot analysis of AKT, p-Akt, mTOR, p-mTOR, FOXO1, and p-FOXO1 expression in U-87 MG cells after treatment with LY294002 (20 μ M), <t>Rapamycin</t> (50 nM), or NAC (5 mM) with or without XHP. (c) Confocal images of U-87 MG cells treated with XHP stained with anti-FOXO1 antibody. (d) Confocal images of U-87 MG cells treated with XHP were stained with anti-p-FOXO1 antibody. (e) Quantitative RT-PCR analysis of U-87 MG cells subjected to FOXO1-specific knockdown, results were normalized with GAPDH level and expressed as folds of control (n = 3, mean ± SD). (f) U-87 MG cells subjected to FOXO1-specific knockdown were assayed by western blot. (g) U-87 MG cells treated with XHP in the absence or presence of LY294002 (20 μ M), Rapamycin (50 nM), or FOXO1-specific siRNA (10 nM). The proportion of Annexin V positive cells was measured by flow cytometry. Data are mean ± SD of three different experiments. ∗P < 0.05, ∗∗P < 0.01 significantly different from the control.
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The <t>Akt/mTOR/FOXO1</t> signaling pathway was associated with the antigliomas of XHP. (a) After being incubated with different doses of XHP, 2 µ M PTX or control medium for 24h, U-87 MG cells were lysed and subjected to western blot analysis of Akt, p-Akt, mTOR, p-mTOR, FOXO1, and p-FOXO1 antibodies. (b) Western blot analysis of AKT, p-Akt, mTOR, p-mTOR, FOXO1, and p-FOXO1 expression in U-87 MG cells after treatment with LY294002 (20 μ M), <t>Rapamycin</t> (50 nM), or NAC (5 mM) with or without XHP. (c) Confocal images of U-87 MG cells treated with XHP stained with anti-FOXO1 antibody. (d) Confocal images of U-87 MG cells treated with XHP were stained with anti-p-FOXO1 antibody. (e) Quantitative RT-PCR analysis of U-87 MG cells subjected to FOXO1-specific knockdown, results were normalized with GAPDH level and expressed as folds of control (n = 3, mean ± SD). (f) U-87 MG cells subjected to FOXO1-specific knockdown were assayed by western blot. (g) U-87 MG cells treated with XHP in the absence or presence of LY294002 (20 μ M), Rapamycin (50 nM), or FOXO1-specific siRNA (10 nM). The proportion of Annexin V positive cells was measured by flow cytometry. Data are mean ± SD of three different experiments. ∗P < 0.05, ∗∗P < 0.01 significantly different from the control.
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The <t>Akt/mTOR/FOXO1</t> signaling pathway was associated with the antigliomas of XHP. (a) After being incubated with different doses of XHP, 2 µ M PTX or control medium for 24h, U-87 MG cells were lysed and subjected to western blot analysis of Akt, p-Akt, mTOR, p-mTOR, FOXO1, and p-FOXO1 antibodies. (b) Western blot analysis of AKT, p-Akt, mTOR, p-mTOR, FOXO1, and p-FOXO1 expression in U-87 MG cells after treatment with LY294002 (20 μ M), <t>Rapamycin</t> (50 nM), or NAC (5 mM) with or without XHP. (c) Confocal images of U-87 MG cells treated with XHP stained with anti-FOXO1 antibody. (d) Confocal images of U-87 MG cells treated with XHP were stained with anti-p-FOXO1 antibody. (e) Quantitative RT-PCR analysis of U-87 MG cells subjected to FOXO1-specific knockdown, results were normalized with GAPDH level and expressed as folds of control (n = 3, mean ± SD). (f) U-87 MG cells subjected to FOXO1-specific knockdown were assayed by western blot. (g) U-87 MG cells treated with XHP in the absence or presence of LY294002 (20 μ M), Rapamycin (50 nM), or FOXO1-specific siRNA (10 nM). The proportion of Annexin V positive cells was measured by flow cytometry. Data are mean ± SD of three different experiments. ∗P < 0.05, ∗∗P < 0.01 significantly different from the control.
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The <t>Akt/mTOR/FOXO1</t> signaling pathway was associated with the antigliomas of XHP. (a) After being incubated with different doses of XHP, 2 µ M PTX or control medium for 24h, U-87 MG cells were lysed and subjected to western blot analysis of Akt, p-Akt, mTOR, p-mTOR, FOXO1, and p-FOXO1 antibodies. (b) Western blot analysis of AKT, p-Akt, mTOR, p-mTOR, FOXO1, and p-FOXO1 expression in U-87 MG cells after treatment with LY294002 (20 μ M), <t>Rapamycin</t> (50 nM), or NAC (5 mM) with or without XHP. (c) Confocal images of U-87 MG cells treated with XHP stained with anti-FOXO1 antibody. (d) Confocal images of U-87 MG cells treated with XHP were stained with anti-p-FOXO1 antibody. (e) Quantitative RT-PCR analysis of U-87 MG cells subjected to FOXO1-specific knockdown, results were normalized with GAPDH level and expressed as folds of control (n = 3, mean ± SD). (f) U-87 MG cells subjected to FOXO1-specific knockdown were assayed by western blot. (g) U-87 MG cells treated with XHP in the absence or presence of LY294002 (20 μ M), Rapamycin (50 nM), or FOXO1-specific siRNA (10 nM). The proportion of Annexin V positive cells was measured by flow cytometry. Data are mean ± SD of three different experiments. ∗P < 0.05, ∗∗P < 0.01 significantly different from the control.
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Image Search Results


The Akt/mTOR/FOXO1 signaling pathway was associated with the antigliomas of XHP. (a) After being incubated with different doses of XHP, 2 µ M PTX or control medium for 24h, U-87 MG cells were lysed and subjected to western blot analysis of Akt, p-Akt, mTOR, p-mTOR, FOXO1, and p-FOXO1 antibodies. (b) Western blot analysis of AKT, p-Akt, mTOR, p-mTOR, FOXO1, and p-FOXO1 expression in U-87 MG cells after treatment with LY294002 (20 μ M), Rapamycin (50 nM), or NAC (5 mM) with or without XHP. (c) Confocal images of U-87 MG cells treated with XHP stained with anti-FOXO1 antibody. (d) Confocal images of U-87 MG cells treated with XHP were stained with anti-p-FOXO1 antibody. (e) Quantitative RT-PCR analysis of U-87 MG cells subjected to FOXO1-specific knockdown, results were normalized with GAPDH level and expressed as folds of control (n = 3, mean ± SD). (f) U-87 MG cells subjected to FOXO1-specific knockdown were assayed by western blot. (g) U-87 MG cells treated with XHP in the absence or presence of LY294002 (20 μ M), Rapamycin (50 nM), or FOXO1-specific siRNA (10 nM). The proportion of Annexin V positive cells was measured by flow cytometry. Data are mean ± SD of three different experiments. ∗P < 0.05, ∗∗P < 0.01 significantly different from the control.

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Xihuang Pill Induces Apoptosis of Human Glioblastoma U-87 MG Cells via Targeting ROS-Mediated Akt/mTOR/FOXO1 Pathway

doi: 10.1155/2018/6049498

Figure Lengend Snippet: The Akt/mTOR/FOXO1 signaling pathway was associated with the antigliomas of XHP. (a) After being incubated with different doses of XHP, 2 µ M PTX or control medium for 24h, U-87 MG cells were lysed and subjected to western blot analysis of Akt, p-Akt, mTOR, p-mTOR, FOXO1, and p-FOXO1 antibodies. (b) Western blot analysis of AKT, p-Akt, mTOR, p-mTOR, FOXO1, and p-FOXO1 expression in U-87 MG cells after treatment with LY294002 (20 μ M), Rapamycin (50 nM), or NAC (5 mM) with or without XHP. (c) Confocal images of U-87 MG cells treated with XHP stained with anti-FOXO1 antibody. (d) Confocal images of U-87 MG cells treated with XHP were stained with anti-p-FOXO1 antibody. (e) Quantitative RT-PCR analysis of U-87 MG cells subjected to FOXO1-specific knockdown, results were normalized with GAPDH level and expressed as folds of control (n = 3, mean ± SD). (f) U-87 MG cells subjected to FOXO1-specific knockdown were assayed by western blot. (g) U-87 MG cells treated with XHP in the absence or presence of LY294002 (20 μ M), Rapamycin (50 nM), or FOXO1-specific siRNA (10 nM). The proportion of Annexin V positive cells was measured by flow cytometry. Data are mean ± SD of three different experiments. ∗P < 0.05, ∗∗P < 0.01 significantly different from the control.

Article Snippet: Mammalian target of rapamycin (mTOR) (2983), phospho-mTOR (Ser2448) (5536), Akt (pan) (4691), phospho-Akt (Ser473) (4060), forkhead transcription factor (FOXO)-1 (C29H4) (2880), phospho-FOXO1 (Thr24) (4G6) (2599), caspase-3 (9662), caspase-9 (9508), B-cell lymphoma-extra large (Bcl-xL) (B9429), Bcl-2-associated X protein (Bax) (5023), and GAPDH (5174) primary antibodies were purchased from Cell Signaling Technology.

Techniques: Incubation, Control, Western Blot, Expressing, Staining, Quantitative RT-PCR, Knockdown, Flow Cytometry