p stat1 701  (Cell Signaling Technology Inc)

Journal: PLoS ONE

Article Title: Altered Protein Networks and Cellular Pathways in Severe West Nile Disease in Mice

doi: 10.1371/journal.pone.0068318

Figure Lengend Snippet: Top three IPA-generated networks of differentially expressed molecules identified by DIGE and iTRAQ labeling between early and mock, late and mock, and late and early WNV-infected brain samples.

Article Snippet: As a positive control for the p-701-STAT1 antibody response, a lysate of murine bone marrow-derived macrophages stimulated for two hours with IFN-γ (PeproTech; 20 ng/ml) was used.

Techniques: Labeling, Cell Function Assay

Journal: PLoS ONE

Article Title: Altered Protein Networks and Cellular Pathways in Severe West Nile Disease in Mice

doi: 10.1371/journal.pone.0068318

Figure Lengend Snippet: (A) Protein samples from each group used for proteomic analysis were minimally labeled with cyanine-3 dye. At the top, a representative protein profile of three biological replicates from brain lysates of mock-, and WNV early- and late-infected mice, separated by 10% SDS-PAGE is shown. WB with fluorescence-based methods was used to detect an overlaid fluorescent scan of the general protein patterns (Cy3 dye (green)) and the specific immunoreactive proteins (FITC or Cy5 dye (red)). To better visualize protein detection signals observed with each specific antibody used, corresponding cropped WB images are presented in grey levels. (B) The graphs correspond to the mean ± S.D. of protein quantity measured by densitometry of the antigenic bands. Densitometry analyses were performed using TotalLab Quant v12.2 software (Nonlinear Dynamics), and data were normalized to levels of global protein pattern intensity. The values indicated under each graph correspond to fold changes from paired comparisons ( i.e., WNV-E/mock, WNV-L/mock, WNV-L/WNV-E). WNV-E/−L, biological replicates of brain samples infected by West Nile Virus and collected at early or late time-points. The significance of the differential protein expression are indicated *, p<0.05; **, p<0.01; ***, p<0.001. A.U., arbitrary units. α-, antibody anti-; ★, IFN-γ activated mice bone marrow derived macrophage (p-701-STAT1 positive control); #, no quantification for p-701-STAT1. CAPN9, calpain 9; CLTC, clathrin heavy chain; DNM1, dynamin 1; GFAP, glial fibrillary acidic protein; HUWE1; E3 ubiquitin-protein ligase; MAP1B, microtubule-associated protein 1B; MAP2, microtubule-associated protein 2; PRDX6, peroxiredoxin 6; STAT1/2, signal transducer and activator of transcription 1/2; VIM, vimentin.

Article Snippet: As a positive control for the p-701-STAT1 antibody response, a lysate of murine bone marrow-derived macrophages stimulated for two hours with IFN-γ (PeproTech; 20 ng/ml) was used.

Techniques: Labeling, Infection, SDS Page, Fluorescence, Software, Expressing, Derivative Assay, Positive Control

Journal: Carcinogenesis

Article Title: Knockdown of ISOC1 inhibits the proliferation and migration and induces the apoptosis of colon cancer cells through the AKT/GSK-3β pathway

doi: 10.1093/carcin/bgz188

Figure Lengend Snippet: The effects of ISOC1 knockdown on the biological function of colon cancer cells are rescued by UCN01. Further verification of the mechanism of ISOC1 in colon cancer cells. The protein level of p-STAT Tyr 701 in colon cancer cell nuclei was increased by ISOC1 knockdown. ( A ) MTS assays. ( B ) Transwell migration assays. ( C ) Cell apoptosis assay. ( D ) Western blot assay. The protein level of p-GSK-3β Ser 9 was upregulated by overexpression of p-AKT Thr 308 . AKT Δ-Thr308 : p-AKT Thr 308 deletion plasmid. AKT P-Thr308 : p-AKT Thr 308 activation plasmid. ( E ) Western blot assay. The protein levels of c-Myc, MMP2 and MMP9 were significantly reduced by ISOC1 knockdown. ( F ) Confocal microscopy and western blot analysis of p-STAT1 Tyr 701 nuclear localization. Green signals represent cells infected with lentiviruses. DNA was visualized with 4′,6-diamidino-2-phenylindole (DAPI) (blue). Western blot analysis showed that the protein levels of p-STAT1 Tyr 701 were increased in the shISOC1 group compared to the shCtrl group. The error bars represent the standard deviation. * P < 0.05, ** P < 0.01.

Article Snippet: The antibodies used for the western blotting included GAPDH (AP0063, Bioworld), ISOC1 (ab118245, Abcam), AKT1/2/3 (ET1609-51, Huabio, China), P-AKT Thr 308 (AF3262, Affinity), P-GSK-3β Ser 9 (AF2016, Affinity), P-STAT1 Tyr 701 (AF6300, Affinity), PARP1 (13371-1-AP, Proteintech), Caspase-3 (9662, CST), Caspase-8 (GB-11594, Servicebio, China), Caspase-9 (GB-11053-1, Servicebio, China), c-Myc (5605s, CST), MMP2 (4022s, CST) and MMP9 (3852s, CST).

Techniques: Knockdown, Migration, Apoptosis Assay, Western Blot, Over Expression, Plasmid Preparation, Activation Assay, Confocal Microscopy, Infection, Standard Deviation