Computational protocol: Enhanced insulin signaling in density-enhanced phosphatase-1 (DEP-1) knockout mice

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Protocol publication

[…] Preparation of protein lysates and wheat germ agglutinin precipitation (for DEP-1 expression analyses) were performed as described in Ref. . Immunoblotting was done by standard protocols with primary antibodies: anti-phospho insulin receptor (IR) Y 972 (ab5678, 1:5000), anti-phospho IR Y 1158 (ab78355, 1:1000), anti-phospho IR Y 1361 (ab60946, 1:1000) (Abcam, Cambridge, UK), anti-DEP-1 (AF1934, 1:1000 of 1 μg/μl dilution, R&D Systems, Wiesbaden, Germany), anti-phospho Akt (#4060, 1:2000, Ser 473), anti-phospho Akt (#9275, 1:2000, Thr 308), anti-pan Akt (#9272, 1:1000) and anti-IR (#3025, 1:1000, 4B8) (Cell Signaling/New England Biolabs, Frankfurt, Germany). Secondary antibodies used were: HRP-linked anti-rabbit (NA934, 1:10,000, GE Healthcare), HRP-linked anti-goat (P 0160, 1:2000, Dako). Densitometric analyses were performed using ImageJ 1.46r. [...] Statistical differences between the groups were determined using two-way ANOVA analysis as well as the non-parametric Mann–Whitney U test and the parametric unpaired Student's t test using SPSS Statistics 21. The data are expressed as means ± SEM, and p < 0.05 was considered statistically significant. […]

Pipeline specifications

Software tools ImageJ, SPSS
Applications Miscellaneous, Microscopic phenotype analysis
Organisms Mus musculus
Diseases Insulin Resistance
Chemicals Glucose