Western Blot Analysis of Lipid Metabolism Proteins

Liver nuclear extracts were prepared as described previously23 (link). Western blotting was performed using specific antibodies against ATGL (Cell Signaling, #2138), mouse CREBH66 (link), HMGCS (Santa Cruz, sc-33828), HSP90 (Santa Cruz, sc-7947), Lamin B1 (Santa Cruz, sc-56145), HSL (Cell Signaling, #4170), phospho-HSL (Ser563) (Cell Signaling, #4139), phospho-HSL (Ser660) (Cell Signaling, #4126), perilipin (Cell Signaling, #9346), and PPARα (Santa Cruz, sc-9000). Following incubation with secondary antibodies, protein bands were visualized by SuperSignal West Pico chemiluminescence substrate (Thermofisher), imaged using Flurochem E system (Proteinsimple, CA, USA), and quantitated using Alphaview software (Proteinsimple, CA, USA).

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Park J.G., Xu X., Cho S., Hur K.Y., Lee M.S., Kersten S, & Lee A.H. (2016). CREBH-FGF21 axis improves hepatic steatosis by suppressing adipose tissue lipolysis. Scientific Reports, 6, 27938.

Publication 2016

HMGCS HSP90 Lamin B1 phospho-HSL (Ser563) phospho-HSL (Ser660) perilipin PPARα SuperSignal West Pico chemiluminescence substrate

Antibodies Chemiluminescence Hsp90 Lamin b1 Liver extracts Mouse Perilipin Protein

Corresponding Organization :

Other organizations : Cornell University, Samsung Medical Center, Sungkyunkwan University, Severance Hospital

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Variable analysis

independent variables

Mouse CREBH
HMGCS
HSP90
Lamin B1
Phospho-HSL (Ser563)
Phospho-HSL (Ser660)
Perilipin
PPARα

dependent variables

Protein bands visualized by SuperSignal West Pico chemiluminescence substrate
Protein bands quantitated using Alphaview software

control variables

Liver nuclear extracts prepared as described previously
Western blotting performed using specific antibodies

controls

Positive control: None specified
Negative control: None specified

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