Protein Expression Analysis in Plantaris Muscle

For protein content analysis, 30 μg of total protein from each plantaris muscle was separated by SDS-PAGE, transferred onto a PVDF membrane, and immunoblotted as previously described [33 (link)]. The following antibodies (Cell Signaling, Danvers, MA) were used: Ulk1 (1:1000), phospho-Ulk1 (1:1000), Drp1 (1:1000), beclin-1 (1:1000), SQSTM1/p62 (1:1000), Bnip3 (1:1000), LC3B (1:1000), and mitofusin-2 (1:1000). Membranes were analyzed and quantified using Bio-Rad Laboratories Image Lab software (Hercules, CA).

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Southern W.M., Nichenko A.S., Shill D.D., Spencer C.C., Jenkins N.T., McCully K.K, & Call J.A. (2017). Skeletal muscle metabolic adaptations to endurance exercise training are attainable in mice with simvastatin treatment. PLoS ONE, 12(2), e0172551.

Publication 2017

phospho-Ulk1 beclin-1 SQSTM1/p62 Bnip3 mitofusin-2 Image Lab software

Antibodies Beclin 1 Membranes Mitofusin 2 Plantaris muscle Protein Pvdf Sds page Ulk1

Corresponding Organization : University of Georgia

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Variable analysis

independent variables

Protein content from plantaris muscle

dependent variables

Levels of Ulk1, phospho-Ulk1, Drp1, beclin-1, SQSTM1/p62, Bnip3, LC3B, and mitofusin-2

control variables

Total protein amount (30 μg) used for SDS-PAGE separation and immunoblotting
PVDF membrane used for protein transfer
Antibody concentrations used for immunoblotting (1:1000 for all antibodies)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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