Protein Expression Quantification in Muscle Biopsies

Protein samples from muscle biopsies were fractionated on SDS–9% PAGE and blotted with anti-β-F1-ATPase (1:1000), anti-Hsp60 (1:1000), anti-GAPDH (1:1000) and anti-PKM2 (1:1000) from [11 (link)], anti-IF1 (1:500) from [12 (link)], anti-LDH-A (1:1000), anti-GPD1 (1:1000) from [13 (link)], anti-PYGM (Abcam, ab88078; 1:1000), anti-PKM1 (Abcam, ab6191-5; 1:1000), anti phospho-PKM2 (Tyr105) (Cell Signaling, #3827; 1:1000), anti-PDHE1α (Invitrogene, 459400; 1:500), anti-phospho-PDHE1α (Ser293) (Abcam, ab92696; 1:1000), anti-PDK1 (Abcam, ab207450; 1:1000) and anti-β-actin (Sigma-Aldrich, A1978; 1:1000). Peroxidase-conjugated anti-mouse IgGs (Nordic Immunology; 1:3000) were used as secondary antibodies. The blots were revealed using the ECL^® reagent (Amersham Pharmacia Biotech). The intensity of the bands was quantified using a Kodak DC120 digital camera and the Kodak 1D Analysis Software.