Journal: Journal of animal science and biotechnology

Article Title: Prepartum body conditions affect insulin signaling pathways in postpartum adipose tissues in transition dairy cows.

doi: 10.1186/s40104-019-0347-4

Figure Lengend Snippet: Fig. 4 Western blot detection of insulin signaling proteins in subcutaneous adipose tissues. These tissues were collected on day 2 postpartum from CON cows (n = 3) and OVE cows (n = 3). a Panels of INSR, IRS1, p-IRS1, AKT, p-AKT (Thr308) and p-AKT (Ser473) protein. β-actin was measured as an internal control. b Intensities of INSR, IRS1, p-IRS1, AKT, p-AKT (Thr308) and p-AKT (Ser473) bands were determined using Quantity One software. The results are presented as the ratio of INSR band intensity to the β-actin band intensity, the ratio of p-ISR1 band intensity to IRS1 band intensity, and the ratio of p-AKT (Thr308) and p-AKT (Ser473) band intensities to the AKT band intensity. IRS1, insulin receptor substrate 1; AKT, protein kinase B

Article Snippet: The following primary antibodies were used: rabbit polyclonal anti-protein kinase B (AKT) (#9272S; 1:1,000 dilution), rabbit monoclonal anti-phospho(p)-AKT (Ser473) (#9272; 1:2,000 dilution), rabbit monoclonal anti-p-AKT (Thr308) (D25E6; 1:1,000 dilution), mouse monoclonal anti-INSR (ab69508; 1:1,000 dilution), rabbit polyclonal anti-insulin receptor substrate 1 (IRS1) (#2382; 1:1,000 dilution), rabbit polyclonal anti-p-IRS1 (#2381; 1:1,000 dilution) (Cell Signaling Technology, Inc., Danvers, MA, USA), and mouse anti-β-actin Mab (66009–1-Ig; 1:1,000 dilution; Proteintech Group Inc., Chicago, IL, USA).

Techniques: Western Blot, Control, Software