p38 mapk isoforms β  (Aviva Systems)

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: p38α MAPK-mediated induction and interaction of FOXO3a and p53 contribute to the inhibited-growth and induced-apoptosis of human lung adenocarcinoma cells by berberine

doi: 10.1186/1756-9966-33-36

Figure Lengend Snippet: Berberine increased the phosphorylation of p38 MAPK and ERK1/2 in a time-dependent manner. A-B , A549 cells were treated with BBR (25 μM) in the indicated times, and cell lysate was harvested and the expression of the phosphorylated or total protein of ERK1/2, p38 MAPK were measured by Western blot analysis using corresponding antibodies. GAPDH was used as loading control. The bar graphs represented the densitometry results of p-ERK (A) or p38 MAPK (B) /GAPDH as mean ± SD of at least three separate experiments. *indicates significant difference from untreated control cells (P < 0.05).

Article Snippet: PD98059 (a special inhibitor of ERK1/2), SB203580 (a special inhibitor of p38 MAPK) were purchased from Merck Millipore (Darmstadt, Germany), MTT powder and Pifithrin-α (PFT-α) were purchased from Sigma Aldrich (St. Louis, MO, USA). p38 MAPK isoforms α and β, p53 and FOXO3a small interfering RNAs (siRNAs) were obtained from Santa Cruz (California, CA, USA).

Techniques: Expressing, Western Blot

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: p38α MAPK-mediated induction and interaction of FOXO3a and p53 contribute to the inhibited-growth and induced-apoptosis of human lung adenocarcinoma cells by berberine

doi: 10.1186/1756-9966-33-36

Figure Lengend Snippet: Berberine increased p53 and FOXO3a protein expression through p38α MAPK pathway. A , A549 cells were exposed to increased concentration of BBR for 24 h. Afterwards, the expression of FOXO3a and p53 protein were detected by Western blot. B - C , A549 cells were treated with SB203580 (10 μM) (B) , or p38α, β siRNAs (70 nM each) (C) for 2 h or 30 h before exposure of the cells to BBR (25 μM) for an additional 24 h. Afterwards, the expression of p38 α or β isoforms, p53 and FOXO3a protein was detected by Western blot. D , A549 cells were treated with PD98059 (20 μM) for 2 h and 30 before exposure of the cells to BBR (25 μM) for an additional 24 h. Afterwards, the expression of p53 and FOXO3a protein was detected by Western blot. The bar graphs represent the mean ± SD of p53/GAPDH and FOXO3a/GAPDH of three independent experiments. E-F , A549 cells were treated with SB203580 (10 μM) for 2 h before exposure of the cells to BBR (25 μM) for an additional 24 h. Afterwards, the cells were collected and processed for analysis of cell cycle distribution by Flow cytometry after propidium iodide (PI) staining (E) . And the percentages of the cell population in each phase (G0/G1, S and G2/M) of cell cycle were assessed by Multicycle AV DNA Analysis Software. Data are expressed as a percentage of total cells. Values are given as the mean ± SD of relative percentage of cell cycle phases from 3 independent experiments performed in triplicate. In separated experiment, the cell viability was determined using the MTT assay (F) . *indicates significant difference as compared to the untreated control group (P < 0.05). **Indicates significant difference from BBR treated alone (P < 0.05).

Article Snippet: PD98059 (a special inhibitor of ERK1/2), SB203580 (a special inhibitor of p38 MAPK) were purchased from Merck Millipore (Darmstadt, Germany), MTT powder and Pifithrin-α (PFT-α) were purchased from Sigma Aldrich (St. Louis, MO, USA). p38 MAPK isoforms α and β, p53 and FOXO3a small interfering RNAs (siRNAs) were obtained from Santa Cruz (California, CA, USA).

Techniques: Expressing, Concentration Assay, Western Blot, Flow Cytometry, Staining, Software, MTT Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: p38α MAPK-mediated induction and interaction of FOXO3a and p53 contribute to the inhibited-growth and induced-apoptosis of human lung adenocarcinoma cells by berberine

doi: 10.1186/1756-9966-33-36

Figure Lengend Snippet: Berberine increased the phosphorylation of p38 MAPK and ERK1/2 in a time-dependent manner. A-B , A549 cells were treated with BBR (25 μM) in the indicated times, and cell lysate was harvested and the expression of the phosphorylated or total protein of ERK1/2, p38 MAPK were measured by Western blot analysis using corresponding antibodies. GAPDH was used as loading control. The bar graphs represented the densitometry results of p-ERK (A) or p38 MAPK (B) /GAPDH as mean ± SD of at least three separate experiments. *indicates significant difference from untreated control cells (P < 0.05).

Article Snippet: Monoclonal antibodies specific for cyclinD1, p38 MAPK isoforms α, extracellular signal-regulated kinase 1/2 (ERK1/2) and their phosphor-forms were purchased from Cell Signaling Technology (Beverly, MA, USA). p38 MAPK isoforms β was ordered from AVIVA System Biology (San Diego, CA, USA).

Techniques: Phospho-proteomics, Expressing, Western Blot, Control

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: p38α MAPK-mediated induction and interaction of FOXO3a and p53 contribute to the inhibited-growth and induced-apoptosis of human lung adenocarcinoma cells by berberine

doi: 10.1186/1756-9966-33-36

Figure Lengend Snippet: Berberine increased p53 and FOXO3a protein expression through p38α MAPK pathway. A , A549 cells were exposed to increased concentration of BBR for 24 h. Afterwards, the expression of FOXO3a and p53 protein were detected by Western blot. B - C , A549 cells were treated with SB203580 (10 μM) (B) , or p38α, β siRNAs (70 nM each) (C) for 2 h or 30 h before exposure of the cells to BBR (25 μM) for an additional 24 h. Afterwards, the expression of p38 α or β isoforms, p53 and FOXO3a protein was detected by Western blot. D , A549 cells were treated with PD98059 (20 μM) for 2 h and 30 before exposure of the cells to BBR (25 μM) for an additional 24 h. Afterwards, the expression of p53 and FOXO3a protein was detected by Western blot. The bar graphs represent the mean ± SD of p53/GAPDH and FOXO3a/GAPDH of three independent experiments. E-F , A549 cells were treated with SB203580 (10 μM) for 2 h before exposure of the cells to BBR (25 μM) for an additional 24 h. Afterwards, the cells were collected and processed for analysis of cell cycle distribution by Flow cytometry after propidium iodide (PI) staining (E) . And the percentages of the cell population in each phase (G0/G1, S and G2/M) of cell cycle were assessed by Multicycle AV DNA Analysis Software. Data are expressed as a percentage of total cells. Values are given as the mean ± SD of relative percentage of cell cycle phases from 3 independent experiments performed in triplicate. In separated experiment, the cell viability was determined using the MTT assay (F) . *indicates significant difference as compared to the untreated control group (P < 0.05). **Indicates significant difference from BBR treated alone (P < 0.05).

Article Snippet: Monoclonal antibodies specific for cyclinD1, p38 MAPK isoforms α, extracellular signal-regulated kinase 1/2 (ERK1/2) and their phosphor-forms were purchased from Cell Signaling Technology (Beverly, MA, USA). p38 MAPK isoforms β was ordered from AVIVA System Biology (San Diego, CA, USA).

Techniques: Expressing, Concentration Assay, Western Blot, Flow Cytometry, Staining, Software, MTT Assay, Control

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: p38α MAPK-mediated induction and interaction of FOXO3a and p53 contribute to the inhibited-growth and induced-apoptosis of human lung adenocarcinoma cells by berberine

doi: 10.1186/1756-9966-33-36

Figure Lengend Snippet: Berberine increased the phosphorylation of p38 MAPK and ERK1/2 in a time-dependent manner. A-B , A549 cells were treated with BBR (25 μM) in the indicated times, and cell lysate was harvested and the expression of the phosphorylated or total protein of ERK1/2, p38 MAPK were measured by Western blot analysis using corresponding antibodies. GAPDH was used as loading control. The bar graphs represented the densitometry results of p-ERK (A) or p38 MAPK (B) /GAPDH as mean ± SD of at least three separate experiments. *indicates significant difference from untreated control cells (P < 0.05).

Article Snippet: Monoclonal antibodies specific for cyclinD1, p38 MAPK isoforms α, extracellular signal-regulated kinase 1/2 (ERK1/2) and their phosphor-forms were purchased from Cell Signaling Technology (Beverly, MA, USA). p38 MAPK isoforms β was ordered from AVIVA System Biology (San Diego, CA, USA).

Techniques: Phospho-proteomics, Expressing, Western Blot, Control

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: p38α MAPK-mediated induction and interaction of FOXO3a and p53 contribute to the inhibited-growth and induced-apoptosis of human lung adenocarcinoma cells by berberine

doi: 10.1186/1756-9966-33-36

Figure Lengend Snippet: Berberine increased p53 and FOXO3a protein expression through p38α MAPK pathway. A , A549 cells were exposed to increased concentration of BBR for 24 h. Afterwards, the expression of FOXO3a and p53 protein were detected by Western blot. B - C , A549 cells were treated with SB203580 (10 μM) (B) , or p38α, β siRNAs (70 nM each) (C) for 2 h or 30 h before exposure of the cells to BBR (25 μM) for an additional 24 h. Afterwards, the expression of p38 α or β isoforms, p53 and FOXO3a protein was detected by Western blot. D , A549 cells were treated with PD98059 (20 μM) for 2 h and 30 before exposure of the cells to BBR (25 μM) for an additional 24 h. Afterwards, the expression of p53 and FOXO3a protein was detected by Western blot. The bar graphs represent the mean ± SD of p53/GAPDH and FOXO3a/GAPDH of three independent experiments. E-F , A549 cells were treated with SB203580 (10 μM) for 2 h before exposure of the cells to BBR (25 μM) for an additional 24 h. Afterwards, the cells were collected and processed for analysis of cell cycle distribution by Flow cytometry after propidium iodide (PI) staining (E) . And the percentages of the cell population in each phase (G0/G1, S and G2/M) of cell cycle were assessed by Multicycle AV DNA Analysis Software. Data are expressed as a percentage of total cells. Values are given as the mean ± SD of relative percentage of cell cycle phases from 3 independent experiments performed in triplicate. In separated experiment, the cell viability was determined using the MTT assay (F) . *indicates significant difference as compared to the untreated control group (P < 0.05). **Indicates significant difference from BBR treated alone (P < 0.05).

Article Snippet: Monoclonal antibodies specific for cyclinD1, p38 MAPK isoforms α, extracellular signal-regulated kinase 1/2 (ERK1/2) and their phosphor-forms were purchased from Cell Signaling Technology (Beverly, MA, USA). p38 MAPK isoforms β was ordered from AVIVA System Biology (San Diego, CA, USA).

Techniques: Expressing, Concentration Assay, Western Blot, Flow Cytometry, Staining, Software, MTT Assay, Control