Journal: Cancer cell international

Article Title: The association between Salt-inducible kinase 2 (SIK2) and gamma isoform of the regulatory subunit B55 of PP2A (B55gamma) contributes to the survival of glioma cells under glucose depletion through inhibiting the phosphorylation of S6K.

doi: 10.1186/s12935-015-0164-6

Figure Lengend Snippet: Figure 3 Overexpression of B55gamma in Glioma cells inhibits the phosphorylation of S6K. A. Overexpression of B55gamma in U251 decreases phosphorylation of S6K, but phosphorylation of 4EBP1 and mTOR and AKT have no significant change. B. U251 cells were transfected with control vector or B55gamma alone or B55gamma + S6K WT or B55gamma + S6K KA for 48 hrs. Cell viability assays were performed under the treatments of Rapamycin at 15 nM and 50 nM, DMSO treatment was control. C. Transfection of S6K WT and S6K KA into U251V and U251B55gamma cells followed by the measurements of cell viabilities under the glucose withdrawal for 24 hrs. Columns, mean of three independent experiments; bars, SE. **, P < 0.01; ***, P < 0.001.

Article Snippet: Vector containing GFPtagged ORF clone of Homo sapiens protein B55gamma, WT S6K or KA S6K were purchased from Origene.

Techniques: Over Expression, Phospho-proteomics, Transfection, Control, Plasmid Preparation

Journal: Cancer cell international

Article Title: The association between Salt-inducible kinase 2 (SIK2) and gamma isoform of the regulatory subunit B55 of PP2A (B55gamma) contributes to the survival of glioma cells under glucose depletion through inhibiting the phosphorylation of S6K.

doi: 10.1186/s12935-015-0164-6

Figure Lengend Snippet: Figure 5 Knockdown of SIK in B55gamma overexpression cells recovers the phosphorylation of S6K. A. U251V, U251B55gamma, U138V and U138B55gamma cells were transfected with scramble siRNA (Ctr) or SIK2 siRNA. 48 hrs after siRNA transfection, cell lysates were prepared and Western blotting was performed to detect the phosphorylation of S6K statues. The β-actin protein was used as a loading control. B. U251B55gamma (left) and U138B55gamma (right) cells were transfected with scramble siRNA (Ctr) or SIK2 siRNA. 48 hrs after siRNA transfection, cell were plated into 6-well plate for overnight, then the regular medium was replaced by glucose free medium for 36 h-starvation followed by the cell viability assay. Columns, mean of three independent experiments; bars, SE. *, P < 0.05.

Article Snippet: Vector containing GFPtagged ORF clone of Homo sapiens protein B55gamma, WT S6K or KA S6K were purchased from Origene.

Techniques: Knockdown, Over Expression, Phospho-proteomics, Transfection, Western Blot, Control, Viability Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Nasal IL-12p70 DNA prevents and treats intestinal allergic diarrhea.

doi: 10.4049/jimmunol.174.11.7423

Figure Lengend Snippet: FIGURE 1. Inhibition of allergic diarrhea disease by nasal treatment with IL-12p70 naked DNA plasmid. A, The incidence of allergic diarrhea was reduced in mice treated nasally with IL-12p70 DNA when compared with mice treated with empty vector plasmid. Allergic disease was induced in these mice by s.c. immunization and then repeated oral challenge (SC/ PO) with OVA. B, OVA-specific IgE Abs are reduced in the serum of allergic diarrhea-afflicted mice treated nasally with IL-12p70 DNA. C, To- tal IgE Abs are reduced in the serum of allergic diarrhea-afflicted mice treated with naked IL-12p70 DNA. The data are expressed as the mean SD and are representative of five independent experiments. Statistical dif- ferences between IL-12p70 DNA- and empty vector-treated mice (, p 0.01 and , p 0.05) are indicated.

Article Snippet: After electrophoresis, proteins were transferred to a polyvinylidene difluoride microporous membrane (Immobilon; Millipore) and the membrane was reacted with biotinylated anti-IL-12p70 (48110.11, rat IgG1; R&D Systems) and then incubated with a biotin-streptavidin complex (ABC-AP kit; Vector Laboratories).

Techniques: Inhibition, Plasmid Preparation

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Nasal IL-12p70 DNA prevents and treats intestinal allergic diarrhea.

doi: 10.4049/jimmunol.174.11.7423

Figure Lengend Snippet: FIGURE 2. Reduction of IgA and IgG hyperresponsiveness in large in- testinal B cells and of Th2 cells responses by nasal administration of IL- 12p70 DNA. A, The frequency of OVA-specific IgA, IgG, IgG1, and IgM AFCs in the large intestine of mice treated with IL-12p70 DNA or empty vector plasmid. B, OVA-specific IL-4 and IFN- production were analyzed by intracellular staining using FACS analysis. In vivo treatment with IL- 12p70 DNA reduced the predominant Th2-type Ag-specific responses by large intestinal mononuclear cells isolated from diarrhea-afflicted mice. The mononuclear cells isolated from the large intestine (1.0 106 cells/ well) or spleen were cultured with OVA (1 mg/ml) for 4 days. After in- cubation, cells were harvested and subjected to intracellular staining with anti-IL-4 and IFN-. The data are expressed as the percentage of cytokine- positive cells in large intestinal CD4 T cells. The data are expressed as the mean SD and are representative of four independent experiments. Sta- tistical differences between IL-12p70 DNA- and empty vector-treated mice (, p 0.01) are indicated. N.D., Not detected.

Article Snippet: After electrophoresis, proteins were transferred to a polyvinylidene difluoride microporous membrane (Immobilon; Millipore) and the membrane was reacted with biotinylated anti-IL-12p70 (48110.11, rat IgG1; R&D Systems) and then incubated with a biotin-streptavidin complex (ABC-AP kit; Vector Laboratories).

Techniques: Plasmid Preparation, Staining, In Vivo, Isolation, Cell Culture

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Nasal IL-12p70 DNA prevents and treats intestinal allergic diarrhea.

doi: 10.4049/jimmunol.174.11.7423

Figure Lengend Snippet: FIGURE 5. Induction of IL-12p70 protein in the large intestine by nasal administration of IL-12p70 DNA. An- ti-IL-12p70 mAb used did not cross- react with the IL-12p40 molecule. A, IL-12p70-positive cells were detected near large intestinal lymphatic vessels after nasal IL-12p70 DNA treatment. B, The IL-12p70 protein was also de- tected in large intestinal extracts using immunoprecipitation and blotting analysis. Recombinant murine IL- 12p70 (rIL-12p70) protein (lane 1) was used as a positive control for the detection of IL-12p70 (B). C, IL- 12p70-producing cells were stained with fluorescence-conjugated anti- CD11c in the large intestine.

Article Snippet: After electrophoresis, proteins were transferred to a polyvinylidene difluoride microporous membrane (Immobilon; Millipore) and the membrane was reacted with biotinylated anti-IL-12p70 (48110.11, rat IgG1; R&D Systems) and then incubated with a biotin-streptavidin complex (ABC-AP kit; Vector Laboratories).

Techniques: Immunoprecipitation, Recombinant, Positive Control, Staining

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Nasal IL-12p70 DNA prevents and treats intestinal allergic diarrhea.

doi: 10.4049/jimmunol.174.11.7423

Figure Lengend Snippet: FIGURE 6. Nasal IL-12p70 DNA treatment cured allergic diarrhea. A, Frequency of allergic diarrhea de- creased after IL-12p70 DNA treat- ment (n 9 per group). B, The symp- toms of diarrhea were completely inhibited after three treatments with IL-12p70 DNA. C, Infiltration of eo- sinophils and basophils into the large intestine was blocked by the IL-12p70 DNA administration. The red arrows point to eosinophils and the blue ar- rows point to basophils. D, Results of the ELISPOT assay used to determine the OVA-specific Ig responses in the large intestinal mononuclear cells. The data are expressed as the mean SD and are representative of three in- dependent experiments. Statistical differences between IL-12p70 DNA- and empty vector-treated mice (, p 0.01) are indicated.

Article Snippet: After electrophoresis, proteins were transferred to a polyvinylidene difluoride microporous membrane (Immobilon; Millipore) and the membrane was reacted with biotinylated anti-IL-12p70 (48110.11, rat IgG1; R&D Systems) and then incubated with a biotin-streptavidin complex (ABC-AP kit; Vector Laboratories).

Techniques: Enzyme-linked Immunospot, Plasmid Preparation