Journal: Antioxidants

Article Title: Intranasal Administration of KYCCSRK Peptide Rescues Brain Insulin Signaling Activation and Reduces Alzheimer’s Disease-like Neuropathology in a Mouse Model for Down Syndrome

doi: 10.3390/antiox12010111

Figure Lengend Snippet: Antibodies used in the current work.

Article Snippet: pIRS1 (S636) , GTX 32400 , Gene Tex , 1:1000.

Techniques:

Journal: Antioxidants

Article Title: Intranasal Administration of KYCCSRK Peptide Rescues Brain Insulin Signaling Activation and Reduces Alzheimer’s Disease-like Neuropathology in a Mouse Model for Down Syndrome

doi: 10.3390/antiox12010111

Figure Lengend Snippet: Activation of insulin signaling following KYCCSRK intranasal administration in Ts2Cje mice. IR, IRS1, AKT, BVR-A, and PTEN protein levels and phosphorylation were evaluated in the frontal cortex of Eu and Ts2Cje (Ts-V) mice treated with vehicle (saline) and Ts2Cje (Ts-P) treated with KYCCSRK peptide (0.5 mM) for two weeks (n = 4/group). ( A ) Representative Western blot and total load images and densitometric evaluation of ( B , C ) IR protein levels and activation [evaluated as pIR Y1146/1150/1151 /IR ratio], ( D ) IRS1 protein levels, ( E ) IRS1 activation evaluated as pIRS1 Y612 /IRS1 ratio], ( F , G ) IRS1 inhibition [evaluated as pIRS1 S636 /IRS1 and pIRS1 S307 /IRS1 ratio], ( H , I ) AKT protein levels and activation (evaluated as pAKT S473 /AKT ratio), ( J ) BVR-A protein levels, and ( K , L ) PTEN protein levels and inhibition (evaluated as pPTEN S380/T382/383 /PTEN). All densitometric values were normalized per total protein load and are given as a percentage of Eu set as 100%. Data are shown as mean ± SEM. One-way ANOVA with Dunnett test: * p < 0.05, ** p < 0.01.

Article Snippet: pIRS1 (S636) , GTX 32400 , Gene Tex , 1:1000.

Techniques: Activation Assay, Phospho-proteomics, Saline, Western Blot, Inhibition

Journal: Molecules and Cells

Article Title: Phosphoserine Phosphatase Promotes Lung Cancer Progression through the Dephosphorylation of IRS-1 and a Noncanonical L-Serine-Independent Pathway

doi: 10.14348/molcells.2019.0160

Figure Lengend Snippet: (A) Validation of the specific interaction between PSPH and IRS-1 proteins. (B) Validation of IRS-1 dephosphorylation by PSPH overexpression (immunoblot assay). (C) Validation of IRS-1 dephosphorylation by wild type (WT) and mutant (D20A) PSPH (immunostaining). Red, phospho-IRS-1 (Ser794); Blue, DAPI. (D) Regulation of downstream signals by WT and mutant (D20A) PSPH. (E) Results of the RT-qPCR assay for the known downstream target genes of the Akt signaling pathway (CDK2: P = 4.0 × 10 −5 , HRAS: P = 7.0 × 10 −5 , MAPKAP1: P = 8.0 × 10 −4 , SLC2A1: P = 1.4 × 10 −6 ; t -test). (F) The correlation between PSPH and Akt target genes. (G) The results of invasion assays with rapamycin treatment after ectopic PSPH overexpression (NCI-H1299: P = 5.8 × 10 −5 , A549: P = 7.4 × 10 −5 ; t -test). Data are representative of three independent experiments. The error bars represent the standard error of the mean. *** P < 0.001.

Article Snippet: For non-small cell lung cancer tissue microarrays (LC121c; US Biomax, USA) staining with PSPH (Sino Biological, China) and pIRS-1 Ser794 (Full Moon BioSystems, USA) antibodies, the antigen retrieval, endogenous peroxidase blocking, primary antibody incubation and DAB development are conducted as same as IHC staining processed.

Techniques: Biomarker Discovery, De-Phosphorylation Assay, Over Expression, Western Blot, Mutagenesis, Immunostaining, Quantitative RT-PCR