primary antibodies against mpc1  (Cell Signaling Technology Inc)

Journal: bioRxiv

Article Title: Sex-dependent adipose glucose partitioning by the mitochondrial pyruvate carrier

doi: 10.1101/2024.05.11.593540

Figure Lengend Snippet: (A - C) Incorporation of [2- 14 C] pyruvate into total lipids ( A ), or the fatty acyl or glycerol-glyceride fraction of lipids ( B – C ) following treatment of differentiated 3T3L1 adipocytes with 5µM UK5099 (A – B) or siRNA against MPC1 or scramble control (C) . (D - E) Total triglyceride accumulation in adipocytes treated with 5µM UK5099 either throughout (D) , or at various time points during (E) , differentiation. (F) Incorporation of [U- 14 C] acetate into total lipids in differentiated 3T3L1 adipocytes treated with or without 5µM UK5099. Data are mean ± SE for at least three experimental replicates (different cell passages). *P<0.05 vs control, ^P<0.05 vs 10% FBS condition in (A) by paired t-test ( B - D and F ), one-way ANOVA ( E ) or 2-way ANOVA ( A ).

Article Snippet: Immunoblot analysis was carried out on cell or adipose lysates using mouse or human primary antibodies against MPC1 (Sigma #HPA045119), MPC2 (Cell Signaling Technologies #46141), GAPDH (Sigma #G8795) and β-Tubulin (Abcam #179513) and developed by chemiluminescence (ECL).

Techniques:

Journal: bioRxiv

Article Title: Sex-dependent adipose glucose partitioning by the mitochondrial pyruvate carrier

doi: 10.1101/2024.05.11.593540

Figure Lengend Snippet: (A - B) Normalized, quantified protein expression and representative western blot of mitochondrial pyruvate carrier proteins in subcutaneous adipose tissue from wild type vs genetically obese (ob/ob) mice ( A ) and from human subjects with normal glucose tolerance vs impaired fasting glucose plus impaired glucose tolerance ( B ). ( C - H ) Pearson correlations between the protein expression of MPC1 ( A , D , E ) or MPC2 ( F , G , H ) in subcutaneous adipose tissue and indices of glucose tolerance ( C and F ), insulin sensitivity ( D and G ) and adipose insulin resistance ( E and H ) in human subjects. Data are n=5 ( A ) or n=7 ( B – H ) in each group. *P<0.05, ***P<0.001 vs control group ( A and B ).

Article Snippet: Immunoblot analysis was carried out on cell or adipose lysates using mouse or human primary antibodies against MPC1 (Sigma #HPA045119), MPC2 (Cell Signaling Technologies #46141), GAPDH (Sigma #G8795) and β-Tubulin (Abcam #179513) and developed by chemiluminescence (ECL).

Techniques: Expressing, Western Blot

Journal: bioRxiv

Article Title: Sex-dependent adipose glucose partitioning by the mitochondrial pyruvate carrier

doi: 10.1101/2024.05.11.593540

Figure Lengend Snippet: (A) MPC1 floxed allele and Adipoq-Cre recombination was visualized by semiquantitative RT-PCR and MPC1 protein knockdown efficacy was verified by western blot of adipose lysates. ( B - C ) Ex vivo incorporation of [2- 14 C] pyruvate into the fatty acyl (lipogenesis B ) or glycerol (glyceroneogenesis C ) moieties of total lipids in epididymal adipose explants from male and female MPC AD-/- mice or LoxP +/+ controls. ( D – E ) Rates of glycerol release ( D ) and non-esterified fatty acid re-esterification ( E ) from epididymal adipose explants from male MPC AD-/- mice or LoxP +/+ controls under basal (unstimulated) conditions or during treatment with insulin or forskolin plus triacsin C. *P<0.05, **P<0.01 for MPC AD-/- vs LoxP +/+ ; ^^P<0.01 for female vs male. Data are mean ± SE for at least five mice per group.

Article Snippet: Immunoblot analysis was carried out on cell or adipose lysates using mouse or human primary antibodies against MPC1 (Sigma #HPA045119), MPC2 (Cell Signaling Technologies #46141), GAPDH (Sigma #G8795) and β-Tubulin (Abcam #179513) and developed by chemiluminescence (ECL).

Techniques: Reverse Transcription Polymerase Chain Reaction, Western Blot, Ex Vivo