Muscle Protein Analysis by Western Blot

Muscle sample preparation and western blot analysis were performed as described previously (26 (link)). The following primary antibodies were used: LC3 (Novus Biological NB100–2220), p62/SQSTM1 (sigma P0067), Atg6 (Novus Biological NB500–249), Atg7 (R&D systems MAB6608), Puromycin (Kerafast EQ001), Akt (Cell signaling CST9272), p-Akt (S473) (Cell signaling CST9271), Raptor (Cell signaling CST2280), p-p70S6k (T389) (Cell signaling CST9205), p70S6k (Abcam AB37699), p-4e-bp-1 (S65) (Cell signaling CST9451), 4e-bp-1 (Cell signaling CST9452), Cox4 (Abcam AB33985), OXPHOS cocktail (Thermo PA5–28220), and Gapdh (Cell signaling CST2118).

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Cui D., Drake J.C., Wilson R.J., Shute R.J., Lewellen B., Zhang M., Zhao H., Sabik O.L., Onengut S., Berr S.S., Rich S.S., Farber C.R, & Yan Z. (2020). A novel voluntary weightlifting model in mice promotes muscle adaptation and insulin sensitivity with simultaneous enhancement of autophagy and mTOR pathway. FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 34(6), 7330-7344.

Publication 2020

p62/SQSTM1 Puromycin p-Akt (S473 Raptor p-p70S6k (T389) p70S6k p-4e-bp-1 (S65 4e-bp-1 OXPHOS cocktail Gapdh

Antibodies Biological Gapdh Muscle Novus P70s6k Puromycin Raptor Western blot analysis

Corresponding Organization : University of Virginia

Other organizations : East China Normal University

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Variable analysis

independent variables

Muscle sample preparation and western blot analysis protocol

dependent variables

Levels of the following proteins: LC3, p62/SQSTM1, Atg6, Atg7, Puromycin, Akt, p-Akt (S473), Raptor, p-p70S6k (T389), p70S6k, p-4e-bp-1 (S65), 4e-bp-1, Cox4, OXPHOS cocktail, Gapdh

control variables

Muscle sample preparation and western blot analysis protocol as described previously (26 (link))

controls

Positive control: None specified
Negative control: None specified

Annotations

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