Muscle Protein Signaling Analysis

Muscle samples were lysed and protein concentration was determined as previously described (26 (link)). Immunoblotting was performed using the following primary antibodies: pAkt Ser⁴⁷³, pAkt Thr³⁰⁸, total Akt, pSAPK/JNK Thr¹⁸³/Tyr¹⁸⁵, total JNK, SIRT1 (mouse specific), acetylated Lys, parkin, LC3, ATG12, PKC-α, PKC-δ, and PKC-θ (Cell Signaling); lipoprotein lipase (LPL) (Abcam); CD36 (R&D Systems); p62 (ProGen); PINK1 (Cayman Chemicals); PGC-1α (Millipore); and α-tubulin (Sigma-Aldrich). For protein kinase C (PKC) studies, subcellular fractionation was carried out as previously described (13 (link)).

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Henstridge D.C., Bruce C.R., Drew B.G., Tory K., Kolonics A., Estevez E., Chung J., Watson N., Gardner T., Lee-Young R.S., Connor T., Watt M.J., Carpenter K., Hargreaves M., McGee S.L., Hevener A.L, & Febbraio M.A. (2014). Activating HSP72 in Rodent Skeletal Muscle Increases Mitochondrial Number and Oxidative Capacity and Decreases Insulin Resistance. Diabetes, 63(6), 1881-1894.

Publication 2014

pAkt Ser473 pAkt Thr308 total Akt pSAPK/JNK Thr183/Tyr185 total JNK parkin ATG12 lipoprotein lipase (LPL) PINK1 PGC-1α α-tubulin

Antibodies Cayman Fractionation Lipoprotein lipase Mouse Muscle Parkin Pgc 1α Progen Protein Protein kinase c Protein kinase c α Protein kinase c δ Protein kinase c θ Sirt1 α tubulin

Corresponding Organization : N-Gene (Hungary)

Other organizations : Monash University, University of California, Los Angeles, Deakin University, University of Sydney, Children's Hospital at Westmead, University of Melbourne

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Variable analysis

independent variables

Protein kinase C (PKC) subcellular fractionation

dependent variables

PAkt Ser^473
PAkt Thr^308
Total Akt
PSAPK/JNK Thr^183/Tyr^185
Total JNK
SIRT1 (mouse specific)
Acetylated Lys
Parkin
ATG12
PKC-α
PKC-δ
PKC-θ
Lipoprotein lipase (LPL)
PINK1
PGC-1α

control variables

Muscle sample lysis and protein concentration determination (as previously described in reference 26)

positive controls

Not specified

negative controls

Not specified

Annotations

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