Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Defining the contribution of skeletal muscle pyruvate dehydrogenase α1 to exercise performance and insulin action

doi: 10.1152/ajpendo.00241.2018

Figure Lengend Snippet: Quantitative PCR primer sequences

Article Snippet: The following antibodies were used: PDHα (cat. no. 3205, Cell Signaling), eukaryotic translation elongation factor 2 (cat. no. 2332, Cell signaling), ATP synthase subunit alpha, ubiquinol-cytochrome C reductase core protein 2, mitochondrially encoded cytochrome C oxidase I, succinate dehydrogenase subunit B, NADH/ubiquinone oxidoreductase subunit B8 (cat. no. MS-604, MitoSciences), acyl-Coenzyme A dehydrogenase, very long-chain; cat. no. ab-155138, Abcam), acyl-Coenzyme A dehydrogenase, long-chain (cat. no. ab-82853, Abcam), hexokinase 2 (HK2; cat. no. 2857, Cell Signaling), lactate dehydrogenase A (cat. no. ABN-896, MilliporeSigma), Akt (cat. no. 2920, Cell Signaling), pAkt S473 (cat. no. 4058, Cell Signaling), GSK3α/β (5676, Cell signaling), pGSK3α/β S21/S9 (9331, Cell signaling), PDK4 (cat. no. NBP1-07047SS, Novus Biologicals), phosphofructokinase 1 (cat. no. sc-377346, Santa Cruz Biotechnology).

Techniques: Real-time Polymerase Chain Reaction

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Defining the contribution of skeletal muscle pyruvate dehydrogenase α1 to exercise performance and insulin action

doi: 10.1152/ajpendo.00241.2018

Figure Lengend Snippet: Metabolic and mitochondrial proteins in HFD-fed PDHmKO and WT mice. WT and PDHmKO mice were fed either a chow or a HFD for 12 wk. Transcript abundance of Pdha1, Pdk2, and Pdk4 (A) and Cd36, Cact (Slc25a20), and Acadl (B) in skeletal muscle from chow- or HFD-fed PDHmKO and WT mice, normalized to Ppib (Chow, n = 5/6; HFD, n = 6/7). Representative blot of PDHα, HK2, LDHA, ACADVL (C), and ATP5A, UQCRC2, MTCO1, SDHB, and NDUFB8 (D) protein abundance in skeletal muscle from chow- or HFD-fed PDHmKO and WT mice. Bar graphs show quantification of protein abundance in skeletal muscle relative to ponceau (Chow, n = 6/6; HFD, n = 6/6) (E). Data reported as means ± SE two-way ANOVA, #P < 0.05, main effect of diet, *P < 0.05, main effect of genotype (A–E). HFD, high-fat diet; PDHmKO, tamoxifen-inducible Pdha1 knockout mice; WT, wild-type.

Article Snippet: The following antibodies were used: PDHα (cat. no. 3205, Cell Signaling), eukaryotic translation elongation factor 2 (cat. no. 2332, Cell signaling), ATP synthase subunit alpha, ubiquinol-cytochrome C reductase core protein 2, mitochondrially encoded cytochrome C oxidase I, succinate dehydrogenase subunit B, NADH/ubiquinone oxidoreductase subunit B8 (cat. no. MS-604, MitoSciences), acyl-Coenzyme A dehydrogenase, very long-chain; cat. no. ab-155138, Abcam), acyl-Coenzyme A dehydrogenase, long-chain (cat. no. ab-82853, Abcam), hexokinase 2 (HK2; cat. no. 2857, Cell Signaling), lactate dehydrogenase A (cat. no. ABN-896, MilliporeSigma), Akt (cat. no. 2920, Cell Signaling), pAkt S473 (cat. no. 4058, Cell Signaling), GSK3α/β (5676, Cell signaling), pGSK3α/β S21/S9 (9331, Cell signaling), PDK4 (cat. no. NBP1-07047SS, Novus Biologicals), phosphofructokinase 1 (cat. no. sc-377346, Santa Cruz Biotechnology).

Techniques: Quantitative Proteomics, Knock-Out

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Defining the contribution of skeletal muscle pyruvate dehydrogenase α1 to exercise performance and insulin action

doi: 10.1152/ajpendo.00241.2018

Figure Lengend Snippet: PDHmKO mice have reduced running speed during voluntary wheel running (VWR). Weekly average speed (km/h) (A), distance run per 24-h period (B), and time/24 h spent running (C) over 21 days VWR (n = 7/6). Representative blots of PDHα, HK2, PFK1, LDH, PDK4, ACADVL, ACADL, and NDUFB8 protein abundance in skeletal muscle from sedentary (SED) or exercise-trained (EXT) PDHmKO and WT mice (D). Bar graphs show quantification of protein abundance in skeletal muscle relative to ponceau (SED, n = 5/5; EXT, n = 7/6) (E). Data reported as means ± SE 2-way ANOVA, Sidak’s post hoc test, *P < 0.05, PDHmKO compared with WT (A–C); 2-way ANOVA, Tukey’s post hoc test, #P < 0.05, EXT compared with SED, *P < 0.05, PDHmKO compared with WT (D–E). PDHmKO, tamoxifen-inducible Pdha1 knockout mice; WT, wild-type.

Article Snippet: The following antibodies were used: PDHα (cat. no. 3205, Cell Signaling), eukaryotic translation elongation factor 2 (cat. no. 2332, Cell signaling), ATP synthase subunit alpha, ubiquinol-cytochrome C reductase core protein 2, mitochondrially encoded cytochrome C oxidase I, succinate dehydrogenase subunit B, NADH/ubiquinone oxidoreductase subunit B8 (cat. no. MS-604, MitoSciences), acyl-Coenzyme A dehydrogenase, very long-chain; cat. no. ab-155138, Abcam), acyl-Coenzyme A dehydrogenase, long-chain (cat. no. ab-82853, Abcam), hexokinase 2 (HK2; cat. no. 2857, Cell Signaling), lactate dehydrogenase A (cat. no. ABN-896, MilliporeSigma), Akt (cat. no. 2920, Cell Signaling), pAkt S473 (cat. no. 4058, Cell Signaling), GSK3α/β (5676, Cell signaling), pGSK3α/β S21/S9 (9331, Cell signaling), PDK4 (cat. no. NBP1-07047SS, Novus Biologicals), phosphofructokinase 1 (cat. no. sc-377346, Santa Cruz Biotechnology).

Techniques: Quantitative Proteomics, Knock-Out

Journal: Clinical and Translational Medicine

Article Title: The antineoplastic drug, trastuzumab, dysregulates metabolism in iPSC-derived cardiomyocytes

doi: 10.1186/s40169-016-0133-2

Figure Lengend Snippet: 76 genes were differentially expressed in both ERBB2-overexpressing cancer cell lines and iPSC-derived cardiomyocytes, following treatment with trastuzumab, p < 0.05

Article Snippet: For detection of PDK4 expression, blots were stained with 2 μg/mL dilution of anti-PDK4 (NBP1-07049, Novus Biologicals).

Techniques:

Journal: Clinical and Translational Medicine

Article Title: The antineoplastic drug, trastuzumab, dysregulates metabolism in iPSC-derived cardiomyocytes

doi: 10.1186/s40169-016-0133-2

Figure Lengend Snippet: Trastuzumab and lapatinib induced down-regulation of PHLDA1 protein expression and up-regulation of PDK4 protein expression. Western blot of 15 μg of total cell lysate of iPSC-derived cardiomyocytes stained for PHLDA1 and PDK4

Article Snippet: For detection of PDK4 expression, blots were stained with 2 μg/mL dilution of anti-PDK4 (NBP1-07049, Novus Biologicals).

Techniques: Expressing, Western Blot, Derivative Assay, Staining