Skeletal Muscle Signaling Pathway Analysis

Skeletal muscle signalling was analysed using methods as previously published^{53 (link)}. The following antibodies were used for the detection of total or phosphorylated protein levels: Hexokinase II (sc 6521), GLUT4 (Abcam 654; samples unboiled), OXPHOS (Abcam 110413), p-Akt Ser473 (CST 9271), p-AMPK (CST 2531), p-ERK 1/2 (CST 4370), p-SEK1/MKK4 (CST 4514), p-P38 (CST 4511), p-JNK (CST 4668), p-SMAD2-L (CST 3104) and p-P70S6K T421/S424 (CST 9204). All primary antibodies were used at 1:1,000 dilution in TBST, with the exception of p-SMAD2-L, which was diluted at 1:500.

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MacDonald T.L., Pattamaprapanont P., Pathak P., Fernandez N., Freitas E.C., Hafida S., Mitri J., Britton S.L., Koch L.G, & Lessard S.J. (2020). Hyperglycaemia is associated with impaired muscle signalling and aerobic adaptation to exercise. Nature metabolism, 2(9), 902-917.

Publication 2020

GLUT4 p-Akt Ser473 p-AMPK p-JNK

Antibodies Dilution Erk 1 Glut4 Hexokinase ii P70s6k Protein Sek1 Skeletal muscle Smad2

Corresponding Organization : Brigham and Women's Hospital

Other organizations : Universidade de São Paulo, University of Michigan–Ann Arbor, University of Toledo

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Total or phosphorylated protein levels of Hexokinase II, GLUT4, OXPHOS, p-Akt Ser473, p-AMPK, p-ERK 1/2, p-SEK1/MKK4, p-P38, p-JNK, p-SMAD2-L, and p-P70S6K T421/S424

control variables

None explicitly mentioned

controls

No positive or negative controls were explicitly mentioned in the input

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