Ab59493

Transfected cells were lysed in RIPA buffer containing 1% phenylmethylsulfonyl fluoride (PMSF). A BCA protein kit (Pierce, Thermo Fisher Scientific Inc., Waltham, MA, USA) was used to determine the total protein concentration. Protein samples were separated using 10% SDS-PAGE and transferred onto PVDF membranes (Millipore, Billerica, MA, USA). The membranes were incubated overnight at 4 °C with the following antibodies: anti-DGAT1 (ab189994, Abcam, Cambridge, UK), anti-DGAT2 (ab59493, Abcam, Cambridge, UK), anti-PPARγ (bs-4509R, Bioss, Beijing, China), anti-C/EBPα (bs-1630R, Bioss, Beijing, China), anti-SREBF1(bs-1402R, Bioss, Beijing, China), anti-Pax7 (ab61067, Abcam, Cambridge, UK), anti-MYOD (bs-2442R, Bioss, Beijing, China), and anti-MYOG (bs-3550R, Bioss, Beijing, China). Thereafter, the membranes were washed and incubated with horseradish peroxidase-conjugated goat anti-mouse or goat anti-rabbit IgG (bs-0295G; Bioss, Beijing, China) for 1 h. β-actin (ab8226, Abcam, Cambridge, UK) was used as an endogenous control. A grayscale intensity analysis was performed using ImageJ software (NIH).

Immunoblotting was performed as described previously (Wiggin et al., 2009 (link)). A total of 20 µg of protein lysate was separated by SDS-PAGE and nitrocellulose membranes were incubated with a goat polyclonal antibody against DGAT2 (Abcam, cat# ab59493, RRID: AB_941282) and a rabbit polyclonal antibody against YWHAZ (Proteintech Group, cat# 14881-1-AP, RRID: AB_2218248).