phospho ikkα β  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho ikkα β
    A & B, Xenograft X1046 cells were disaggregated into single cells and briefly propagated as neurospheres in vitro . Cells were pre-treated with AZD1480 (1 µM) and/or WA (5 µM) for 2 h prior to TNF-α (10 ng/ml) for 0.25 or 2 h (A) or 2 h (B). Cells were lysed and immunoblotted with the indicated Ab (A), or RNA was isolated followed by generation of cDNA and qRT-PCR was performed for IL-6 (B). Densitometric values of p-p65, <t>p-IKKα/β</t> and p-STAT3 were normalized to total p65, total IKKα/β and total STAT3, respectively. Data are shown as replicates of three. *, p<0.05. C, Xenograft X1066 cells were treated with the indicated doses of AZD1480 and/or WA for 48 h, and the WST-1 cell viability assay was performed. Data are shown as replicates of three. *p<0.05. D, Nude mice were injected intracranially with Xenograft X1016 cells. Starting at day 3, mice were treated with vehicle (n = 5), AZD1480 (30 mg/kg, twice a day, n = 5), WA (4 mg/kg, alternate days, n = 5) or both AZD+WA (n = 5) for three weeks. Survival was measured, and mice were euthanized at moribund. *, p<0.05 (LogRank).
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    1) Product Images from "NF-κB-Induced IL-6 Ensures STAT3 Activation and Tumor Aggressiveness in Glioblastoma"

    Article Title: NF-κB-Induced IL-6 Ensures STAT3 Activation and Tumor Aggressiveness in Glioblastoma

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0078728

    A & B, Xenograft X1046 cells were disaggregated into single cells and briefly propagated as neurospheres in vitro . Cells were pre-treated with AZD1480 (1 µM) and/or WA (5 µM) for 2 h prior to TNF-α (10 ng/ml) for 0.25 or 2 h (A) or 2 h (B). Cells were lysed and immunoblotted with the indicated Ab (A), or RNA was isolated followed by generation of cDNA and qRT-PCR was performed for IL-6 (B). Densitometric values of p-p65, p-IKKα/β and p-STAT3 were normalized to total p65, total IKKα/β and total STAT3, respectively. Data are shown as replicates of three. *, p<0.05. C, Xenograft X1066 cells were treated with the indicated doses of AZD1480 and/or WA for 48 h, and the WST-1 cell viability assay was performed. Data are shown as replicates of three. *p<0.05. D, Nude mice were injected intracranially with Xenograft X1016 cells. Starting at day 3, mice were treated with vehicle (n = 5), AZD1480 (30 mg/kg, twice a day, n = 5), WA (4 mg/kg, alternate days, n = 5) or both AZD+WA (n = 5) for three weeks. Survival was measured, and mice were euthanized at moribund. *, p<0.05 (LogRank).
    Figure Legend Snippet: A & B, Xenograft X1046 cells were disaggregated into single cells and briefly propagated as neurospheres in vitro . Cells were pre-treated with AZD1480 (1 µM) and/or WA (5 µM) for 2 h prior to TNF-α (10 ng/ml) for 0.25 or 2 h (A) or 2 h (B). Cells were lysed and immunoblotted with the indicated Ab (A), or RNA was isolated followed by generation of cDNA and qRT-PCR was performed for IL-6 (B). Densitometric values of p-p65, p-IKKα/β and p-STAT3 were normalized to total p65, total IKKα/β and total STAT3, respectively. Data are shown as replicates of three. *, p<0.05. C, Xenograft X1066 cells were treated with the indicated doses of AZD1480 and/or WA for 48 h, and the WST-1 cell viability assay was performed. Data are shown as replicates of three. *p<0.05. D, Nude mice were injected intracranially with Xenograft X1016 cells. Starting at day 3, mice were treated with vehicle (n = 5), AZD1480 (30 mg/kg, twice a day, n = 5), WA (4 mg/kg, alternate days, n = 5) or both AZD+WA (n = 5) for three weeks. Survival was measured, and mice were euthanized at moribund. *, p<0.05 (LogRank).

    Techniques Used: In Vitro, Isolation, Quantitative RT-PCR, Viability Assay, Injection

    phospho ikkα β  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho ikkα β
    A & B, Xenograft X1046 cells were disaggregated into single cells and briefly propagated as neurospheres in vitro . Cells were pre-treated with AZD1480 (1 µM) and/or WA (5 µM) for 2 h prior to TNF-α (10 ng/ml) for 0.25 or 2 h (A) or 2 h (B). Cells were lysed and immunoblotted with the indicated Ab (A), or RNA was isolated followed by generation of cDNA and qRT-PCR was performed for IL-6 (B). Densitometric values of p-p65, <t>p-IKKα/β</t> and p-STAT3 were normalized to total p65, total IKKα/β and total STAT3, respectively. Data are shown as replicates of three. *, p<0.05. C, Xenograft X1066 cells were treated with the indicated doses of AZD1480 and/or WA for 48 h, and the WST-1 cell viability assay was performed. Data are shown as replicates of three. *p<0.05. D, Nude mice were injected intracranially with Xenograft X1016 cells. Starting at day 3, mice were treated with vehicle (n = 5), AZD1480 (30 mg/kg, twice a day, n = 5), WA (4 mg/kg, alternate days, n = 5) or both AZD+WA (n = 5) for three weeks. Survival was measured, and mice were euthanized at moribund. *, p<0.05 (LogRank).
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    1) Product Images from "NF-κB-Induced IL-6 Ensures STAT3 Activation and Tumor Aggressiveness in Glioblastoma"

    Article Title: NF-κB-Induced IL-6 Ensures STAT3 Activation and Tumor Aggressiveness in Glioblastoma

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0078728

    A & B, Xenograft X1046 cells were disaggregated into single cells and briefly propagated as neurospheres in vitro . Cells were pre-treated with AZD1480 (1 µM) and/or WA (5 µM) for 2 h prior to TNF-α (10 ng/ml) for 0.25 or 2 h (A) or 2 h (B). Cells were lysed and immunoblotted with the indicated Ab (A), or RNA was isolated followed by generation of cDNA and qRT-PCR was performed for IL-6 (B). Densitometric values of p-p65, p-IKKα/β and p-STAT3 were normalized to total p65, total IKKα/β and total STAT3, respectively. Data are shown as replicates of three. *, p<0.05. C, Xenograft X1066 cells were treated with the indicated doses of AZD1480 and/or WA for 48 h, and the WST-1 cell viability assay was performed. Data are shown as replicates of three. *p<0.05. D, Nude mice were injected intracranially with Xenograft X1016 cells. Starting at day 3, mice were treated with vehicle (n = 5), AZD1480 (30 mg/kg, twice a day, n = 5), WA (4 mg/kg, alternate days, n = 5) or both AZD+WA (n = 5) for three weeks. Survival was measured, and mice were euthanized at moribund. *, p<0.05 (LogRank).
    Figure Legend Snippet: A & B, Xenograft X1046 cells were disaggregated into single cells and briefly propagated as neurospheres in vitro . Cells were pre-treated with AZD1480 (1 µM) and/or WA (5 µM) for 2 h prior to TNF-α (10 ng/ml) for 0.25 or 2 h (A) or 2 h (B). Cells were lysed and immunoblotted with the indicated Ab (A), or RNA was isolated followed by generation of cDNA and qRT-PCR was performed for IL-6 (B). Densitometric values of p-p65, p-IKKα/β and p-STAT3 were normalized to total p65, total IKKα/β and total STAT3, respectively. Data are shown as replicates of three. *, p<0.05. C, Xenograft X1066 cells were treated with the indicated doses of AZD1480 and/or WA for 48 h, and the WST-1 cell viability assay was performed. Data are shown as replicates of three. *p<0.05. D, Nude mice were injected intracranially with Xenograft X1016 cells. Starting at day 3, mice were treated with vehicle (n = 5), AZD1480 (30 mg/kg, twice a day, n = 5), WA (4 mg/kg, alternate days, n = 5) or both AZD+WA (n = 5) for three weeks. Survival was measured, and mice were euthanized at moribund. *, p<0.05 (LogRank).

    Techniques Used: In Vitro, Isolation, Quantitative RT-PCR, Viability Assay, Injection

    phospho akt473  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho akt473
    Phospho Akt473, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    phospho akt308  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho akt308
    K562 cells with or without ME2 knockdown were lysed with RIPA lysis buffer (50 mM Tris-HCl, pH 8.0, 150 mM NaCl, 1% Triton X-100, 1 mM EGTA) containing 1 mM PMSF and a protease inhibitor cocktail and subjected to centrifugation at 15,000× g for 10 min at 4°C to remove debris. After lysis, equal aliquots of protein lysate were resolved by Western blotting. Western blots were probed with anti-phospho-ERK1/2, anti-ERK1, <t>anti-p-AKT308,</t> anti-AKT472, anti-AKT1/2, anti-GATA-1, anti-vimentin and anti-β-tubulin. A, phospho-ERK1/2 activity in ME2 knockdown K562 cells. B, Phospho-AKT detection in ME2 knockdown K562 cells. C, 10 µM PI3K inhibitor, LY294002, inhibits p-AKT activity. D, LY294002 rescue of differentiation in ME2 knockdown K562 cells. E, The effect of LY294002 on the proliferation of K562 cells with or without ME2 knockdown. F, The expression difference of GATA-1 and vimentin in ME2 knockdown cells. Data are representative of three independent experiments.
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    1) Product Images from "Induction of Erythroid Differentiation in Human Erythroleukemia Cells by Depletion of Malic Enzyme 2"

    Article Title: Induction of Erythroid Differentiation in Human Erythroleukemia Cells by Depletion of Malic Enzyme 2

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0012520

    K562 cells with or without ME2 knockdown were lysed with RIPA lysis buffer (50 mM Tris-HCl, pH 8.0, 150 mM NaCl, 1% Triton X-100, 1 mM EGTA) containing 1 mM PMSF and a protease inhibitor cocktail and subjected to centrifugation at 15,000× g for 10 min at 4°C to remove debris. After lysis, equal aliquots of protein lysate were resolved by Western blotting. Western blots were probed with anti-phospho-ERK1/2, anti-ERK1, anti-p-AKT308, anti-AKT472, anti-AKT1/2, anti-GATA-1, anti-vimentin and anti-β-tubulin. A, phospho-ERK1/2 activity in ME2 knockdown K562 cells. B, Phospho-AKT detection in ME2 knockdown K562 cells. C, 10 µM PI3K inhibitor, LY294002, inhibits p-AKT activity. D, LY294002 rescue of differentiation in ME2 knockdown K562 cells. E, The effect of LY294002 on the proliferation of K562 cells with or without ME2 knockdown. F, The expression difference of GATA-1 and vimentin in ME2 knockdown cells. Data are representative of three independent experiments.
    Figure Legend Snippet: K562 cells with or without ME2 knockdown were lysed with RIPA lysis buffer (50 mM Tris-HCl, pH 8.0, 150 mM NaCl, 1% Triton X-100, 1 mM EGTA) containing 1 mM PMSF and a protease inhibitor cocktail and subjected to centrifugation at 15,000× g for 10 min at 4°C to remove debris. After lysis, equal aliquots of protein lysate were resolved by Western blotting. Western blots were probed with anti-phospho-ERK1/2, anti-ERK1, anti-p-AKT308, anti-AKT472, anti-AKT1/2, anti-GATA-1, anti-vimentin and anti-β-tubulin. A, phospho-ERK1/2 activity in ME2 knockdown K562 cells. B, Phospho-AKT detection in ME2 knockdown K562 cells. C, 10 µM PI3K inhibitor, LY294002, inhibits p-AKT activity. D, LY294002 rescue of differentiation in ME2 knockdown K562 cells. E, The effect of LY294002 on the proliferation of K562 cells with or without ME2 knockdown. F, The expression difference of GATA-1 and vimentin in ME2 knockdown cells. Data are representative of three independent experiments.

    Techniques Used: Lysis, Protease Inhibitor, Centrifugation, Western Blot, Activity Assay, Expressing

    anti phospho iκb  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti phospho iκb
    Anti Phospho Iκb, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    total mapkinase  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc total mapkinase
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    phospho  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho
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    polyclonal anti mouse  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc polyclonal anti mouse
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    phospho pkb  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho pkb
    Insulin sensitivity is maintained during anaesthesia if hypothermia is prevented . Mice were injected i.p. with ketamine/Xylazine (K/X), and body temperature maintained at 37°C. Half of the group (n = 4) were given insulin by icv injection, the other half (n = 4) received saline 30 mins prior to sacrifice and rapid removal and dissection of the brain. Equal amounts of cortical protein were subjected to western blot using <t>the</t> <t>antibodies</t> shown (A), and results from each treatment group quantified by densitometry (B). The mean ± SD, relative to the control animals, is shown. Significant differences between control and each treatment group are given, *p < 0.05, **p < 0.005. Phosphorylation of ERK and <t>PKB</t> was also compared between animals sacrificed without anaesthesia and those with anaesthesia but whose body temperature was maintained at 37°C (C).
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    1) Product Images from "Anaesthesia generates neuronal insulin resistance by inducing hypothermia"

    Article Title: Anaesthesia generates neuronal insulin resistance by inducing hypothermia

    Journal: BMC Neuroscience

    doi: 10.1186/1471-2202-9-100

    Insulin sensitivity is maintained during anaesthesia if hypothermia is prevented . Mice were injected i.p. with ketamine/Xylazine (K/X), and body temperature maintained at 37°C. Half of the group (n = 4) were given insulin by icv injection, the other half (n = 4) received saline 30 mins prior to sacrifice and rapid removal and dissection of the brain. Equal amounts of cortical protein were subjected to western blot using the antibodies shown (A), and results from each treatment group quantified by densitometry (B). The mean ± SD, relative to the control animals, is shown. Significant differences between control and each treatment group are given, *p < 0.05, **p < 0.005. Phosphorylation of ERK and PKB was also compared between animals sacrificed without anaesthesia and those with anaesthesia but whose body temperature was maintained at 37°C (C).
    Figure Legend Snippet: Insulin sensitivity is maintained during anaesthesia if hypothermia is prevented . Mice were injected i.p. with ketamine/Xylazine (K/X), and body temperature maintained at 37°C. Half of the group (n = 4) were given insulin by icv injection, the other half (n = 4) received saline 30 mins prior to sacrifice and rapid removal and dissection of the brain. Equal amounts of cortical protein were subjected to western blot using the antibodies shown (A), and results from each treatment group quantified by densitometry (B). The mean ± SD, relative to the control animals, is shown. Significant differences between control and each treatment group are given, *p < 0.05, **p < 0.005. Phosphorylation of ERK and PKB was also compared between animals sacrificed without anaesthesia and those with anaesthesia but whose body temperature was maintained at 37°C (C).

    Techniques Used: Injection, Dissection, Western Blot

    anti bodies  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti bodies
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    anti phospho ampkß s108  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti phospho ampkß s108
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    Cell Signaling Technology Inc phospho ikkα β
    A & B, Xenograft X1046 cells were disaggregated into single cells and briefly propagated as neurospheres in vitro . Cells were pre-treated with AZD1480 (1 µM) and/or WA (5 µM) for 2 h prior to TNF-α (10 ng/ml) for 0.25 or 2 h (A) or 2 h (B). Cells were lysed and immunoblotted with the indicated Ab (A), or RNA was isolated followed by generation of cDNA and qRT-PCR was performed for IL-6 (B). Densitometric values of p-p65, <t>p-IKKα/β</t> and p-STAT3 were normalized to total p65, total IKKα/β and total STAT3, respectively. Data are shown as replicates of three. *, p<0.05. C, Xenograft X1066 cells were treated with the indicated doses of AZD1480 and/or WA for 48 h, and the WST-1 cell viability assay was performed. Data are shown as replicates of three. *p<0.05. D, Nude mice were injected intracranially with Xenograft X1016 cells. Starting at day 3, mice were treated with vehicle (n = 5), AZD1480 (30 mg/kg, twice a day, n = 5), WA (4 mg/kg, alternate days, n = 5) or both AZD+WA (n = 5) for three weeks. Survival was measured, and mice were euthanized at moribund. *, p<0.05 (LogRank).
    Phospho Ikkα β, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc phospho akt473
    A & B, Xenograft X1046 cells were disaggregated into single cells and briefly propagated as neurospheres in vitro . Cells were pre-treated with AZD1480 (1 µM) and/or WA (5 µM) for 2 h prior to TNF-α (10 ng/ml) for 0.25 or 2 h (A) or 2 h (B). Cells were lysed and immunoblotted with the indicated Ab (A), or RNA was isolated followed by generation of cDNA and qRT-PCR was performed for IL-6 (B). Densitometric values of p-p65, <t>p-IKKα/β</t> and p-STAT3 were normalized to total p65, total IKKα/β and total STAT3, respectively. Data are shown as replicates of three. *, p<0.05. C, Xenograft X1066 cells were treated with the indicated doses of AZD1480 and/or WA for 48 h, and the WST-1 cell viability assay was performed. Data are shown as replicates of three. *p<0.05. D, Nude mice were injected intracranially with Xenograft X1016 cells. Starting at day 3, mice were treated with vehicle (n = 5), AZD1480 (30 mg/kg, twice a day, n = 5), WA (4 mg/kg, alternate days, n = 5) or both AZD+WA (n = 5) for three weeks. Survival was measured, and mice were euthanized at moribund. *, p<0.05 (LogRank).
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    Cell Signaling Technology Inc phospho akt308
    K562 cells with or without ME2 knockdown were lysed with RIPA lysis buffer (50 mM Tris-HCl, pH 8.0, 150 mM NaCl, 1% Triton X-100, 1 mM EGTA) containing 1 mM PMSF and a protease inhibitor cocktail and subjected to centrifugation at 15,000× g for 10 min at 4°C to remove debris. After lysis, equal aliquots of protein lysate were resolved by Western blotting. Western blots were probed with anti-phospho-ERK1/2, anti-ERK1, <t>anti-p-AKT308,</t> anti-AKT472, anti-AKT1/2, anti-GATA-1, anti-vimentin and anti-β-tubulin. A, phospho-ERK1/2 activity in ME2 knockdown K562 cells. B, Phospho-AKT detection in ME2 knockdown K562 cells. C, 10 µM PI3K inhibitor, LY294002, inhibits p-AKT activity. D, LY294002 rescue of differentiation in ME2 knockdown K562 cells. E, The effect of LY294002 on the proliferation of K562 cells with or without ME2 knockdown. F, The expression difference of GATA-1 and vimentin in ME2 knockdown cells. Data are representative of three independent experiments.
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    K562 cells with or without ME2 knockdown were lysed with RIPA lysis buffer (50 mM Tris-HCl, pH 8.0, 150 mM NaCl, 1% Triton X-100, 1 mM EGTA) containing 1 mM PMSF and a protease inhibitor cocktail and subjected to centrifugation at 15,000× g for 10 min at 4°C to remove debris. After lysis, equal aliquots of protein lysate were resolved by Western blotting. Western blots were probed with anti-phospho-ERK1/2, anti-ERK1, <t>anti-p-AKT308,</t> anti-AKT472, anti-AKT1/2, anti-GATA-1, anti-vimentin and anti-β-tubulin. A, phospho-ERK1/2 activity in ME2 knockdown K562 cells. B, Phospho-AKT detection in ME2 knockdown K562 cells. C, 10 µM PI3K inhibitor, LY294002, inhibits p-AKT activity. D, LY294002 rescue of differentiation in ME2 knockdown K562 cells. E, The effect of LY294002 on the proliferation of K562 cells with or without ME2 knockdown. F, The expression difference of GATA-1 and vimentin in ME2 knockdown cells. Data are representative of three independent experiments.
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    Image Search Results


    A & B, Xenograft X1046 cells were disaggregated into single cells and briefly propagated as neurospheres in vitro . Cells were pre-treated with AZD1480 (1 µM) and/or WA (5 µM) for 2 h prior to TNF-α (10 ng/ml) for 0.25 or 2 h (A) or 2 h (B). Cells were lysed and immunoblotted with the indicated Ab (A), or RNA was isolated followed by generation of cDNA and qRT-PCR was performed for IL-6 (B). Densitometric values of p-p65, p-IKKα/β and p-STAT3 were normalized to total p65, total IKKα/β and total STAT3, respectively. Data are shown as replicates of three. *, p<0.05. C, Xenograft X1066 cells were treated with the indicated doses of AZD1480 and/or WA for 48 h, and the WST-1 cell viability assay was performed. Data are shown as replicates of three. *p<0.05. D, Nude mice were injected intracranially with Xenograft X1016 cells. Starting at day 3, mice were treated with vehicle (n = 5), AZD1480 (30 mg/kg, twice a day, n = 5), WA (4 mg/kg, alternate days, n = 5) or both AZD+WA (n = 5) for three weeks. Survival was measured, and mice were euthanized at moribund. *, p<0.05 (LogRank).

    Journal: PLoS ONE

    Article Title: NF-κB-Induced IL-6 Ensures STAT3 Activation and Tumor Aggressiveness in Glioblastoma

    doi: 10.1371/journal.pone.0078728

    Figure Lengend Snippet: A & B, Xenograft X1046 cells were disaggregated into single cells and briefly propagated as neurospheres in vitro . Cells were pre-treated with AZD1480 (1 µM) and/or WA (5 µM) for 2 h prior to TNF-α (10 ng/ml) for 0.25 or 2 h (A) or 2 h (B). Cells were lysed and immunoblotted with the indicated Ab (A), or RNA was isolated followed by generation of cDNA and qRT-PCR was performed for IL-6 (B). Densitometric values of p-p65, p-IKKα/β and p-STAT3 were normalized to total p65, total IKKα/β and total STAT3, respectively. Data are shown as replicates of three. *, p<0.05. C, Xenograft X1066 cells were treated with the indicated doses of AZD1480 and/or WA for 48 h, and the WST-1 cell viability assay was performed. Data are shown as replicates of three. *p<0.05. D, Nude mice were injected intracranially with Xenograft X1016 cells. Starting at day 3, mice were treated with vehicle (n = 5), AZD1480 (30 mg/kg, twice a day, n = 5), WA (4 mg/kg, alternate days, n = 5) or both AZD+WA (n = 5) for three weeks. Survival was measured, and mice were euthanized at moribund. *, p<0.05 (LogRank).

    Article Snippet: Antibodies against phospho-p65 Ser536 (p-p65 (S536)), phospho-STAT3 Tyr705 (p-STAT3 (Y705)), STAT3, phospho-IKKα/β (S176/180) and IKKα/β were purchased from Cell Signaling Technology (1∶1,000; Beverly, MA).

    Techniques: In Vitro, Isolation, Quantitative RT-PCR, Viability Assay, Injection

    K562 cells with or without ME2 knockdown were lysed with RIPA lysis buffer (50 mM Tris-HCl, pH 8.0, 150 mM NaCl, 1% Triton X-100, 1 mM EGTA) containing 1 mM PMSF and a protease inhibitor cocktail and subjected to centrifugation at 15,000× g for 10 min at 4°C to remove debris. After lysis, equal aliquots of protein lysate were resolved by Western blotting. Western blots were probed with anti-phospho-ERK1/2, anti-ERK1, anti-p-AKT308, anti-AKT472, anti-AKT1/2, anti-GATA-1, anti-vimentin and anti-β-tubulin. A, phospho-ERK1/2 activity in ME2 knockdown K562 cells. B, Phospho-AKT detection in ME2 knockdown K562 cells. C, 10 µM PI3K inhibitor, LY294002, inhibits p-AKT activity. D, LY294002 rescue of differentiation in ME2 knockdown K562 cells. E, The effect of LY294002 on the proliferation of K562 cells with or without ME2 knockdown. F, The expression difference of GATA-1 and vimentin in ME2 knockdown cells. Data are representative of three independent experiments.

    Journal: PLoS ONE

    Article Title: Induction of Erythroid Differentiation in Human Erythroleukemia Cells by Depletion of Malic Enzyme 2

    doi: 10.1371/journal.pone.0012520

    Figure Lengend Snippet: K562 cells with or without ME2 knockdown were lysed with RIPA lysis buffer (50 mM Tris-HCl, pH 8.0, 150 mM NaCl, 1% Triton X-100, 1 mM EGTA) containing 1 mM PMSF and a protease inhibitor cocktail and subjected to centrifugation at 15,000× g for 10 min at 4°C to remove debris. After lysis, equal aliquots of protein lysate were resolved by Western blotting. Western blots were probed with anti-phospho-ERK1/2, anti-ERK1, anti-p-AKT308, anti-AKT472, anti-AKT1/2, anti-GATA-1, anti-vimentin and anti-β-tubulin. A, phospho-ERK1/2 activity in ME2 knockdown K562 cells. B, Phospho-AKT detection in ME2 knockdown K562 cells. C, 10 µM PI3K inhibitor, LY294002, inhibits p-AKT activity. D, LY294002 rescue of differentiation in ME2 knockdown K562 cells. E, The effect of LY294002 on the proliferation of K562 cells with or without ME2 knockdown. F, The expression difference of GATA-1 and vimentin in ME2 knockdown cells. Data are representative of three independent experiments.

    Article Snippet: The anti-GATA-1, phospho-ERK1/2, phospho-AKT308, phospho-AKT473 and AKT1/2 polyclonal antibodies were purchased from Cell Signaling Technology.

    Techniques: Lysis, Protease Inhibitor, Centrifugation, Western Blot, Activity Assay, Expressing

    Insulin sensitivity is maintained during anaesthesia if hypothermia is prevented . Mice were injected i.p. with ketamine/Xylazine (K/X), and body temperature maintained at 37°C. Half of the group (n = 4) were given insulin by icv injection, the other half (n = 4) received saline 30 mins prior to sacrifice and rapid removal and dissection of the brain. Equal amounts of cortical protein were subjected to western blot using the antibodies shown (A), and results from each treatment group quantified by densitometry (B). The mean ± SD, relative to the control animals, is shown. Significant differences between control and each treatment group are given, *p < 0.05, **p < 0.005. Phosphorylation of ERK and PKB was also compared between animals sacrificed without anaesthesia and those with anaesthesia but whose body temperature was maintained at 37°C (C).

    Journal: BMC Neuroscience

    Article Title: Anaesthesia generates neuronal insulin resistance by inducing hypothermia

    doi: 10.1186/1471-2202-9-100

    Figure Lengend Snippet: Insulin sensitivity is maintained during anaesthesia if hypothermia is prevented . Mice were injected i.p. with ketamine/Xylazine (K/X), and body temperature maintained at 37°C. Half of the group (n = 4) were given insulin by icv injection, the other half (n = 4) received saline 30 mins prior to sacrifice and rapid removal and dissection of the brain. Equal amounts of cortical protein were subjected to western blot using the antibodies shown (A), and results from each treatment group quantified by densitometry (B). The mean ± SD, relative to the control animals, is shown. Significant differences between control and each treatment group are given, *p < 0.05, **p < 0.005. Phosphorylation of ERK and PKB was also compared between animals sacrificed without anaesthesia and those with anaesthesia but whose body temperature was maintained at 37°C (C).

    Article Snippet: Antibodies to phospho-PKB (Thr308), phospho-PKB (Ser473), phospho-ERK (Thr202/Tyr204), were from Cell Signaling Technology (Beverly, MA, U.S.A.), antibodies to total PKB, ERK1/2 and GSK3α/β were from Upstate Biotechnologies (Lake Placid, NY, U.S.A.).

    Techniques: Injection, Dissection, Western Blot