antibodies against mpc1  (Novus Biologicals)

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

Journal: Molecular Cancer

Article Title: 3-BrPA eliminates human bladder cancer cells with highly oncogenic signatures via engagement of specific death programs and perturbation of multiple signaling and metabolic determinants

doi: 10.1186/s12943-015-0399-9

Figure Lengend Snippet: MCT family-member and macropinocytosis activities critically regulate bladder cancer cell susceptibility to 3-BrPA - the dispensable role of MPC components. ( a ) Representative (three independent experiments) Western blotting profiles of whole-cell protein extracts obtained from RT4 and T24 cells, seeded at ~60 % confluency and treated with the indicated doses of 3-BrPA for 24 h. The proteins examined were MCT1, MCT4, SMCT1, MPC1 and MPC2, while Actin was used as molecule of reference. ( b ) Representative (three independent experiments) immunofluorescence images of MCT1 expression and localization in RT4 and T24 cells, seeded at ~60 % confluency and exposed to the indicated doses of 3-BrPA for 4 h (also, see Fig. ). Scale bars: 3 μm. ( c - e ) MTT cytotoxicity assays of T24 cells, grown at low (and ~60 %; data not shown) confluency and treated with the indicated doses of 3-BrPA for 24 h, in the absence or presence (pre-incubation for the denoted time) of 1 or 10 μM UK-5099 ( c ), 0.1 or 1 μM AR-C155858 ( d ) and 10 or 50 μM EIPA ( e ) (both low and ~60 % confluency allowed the striking survival of 3-BrPA-treated T24 cells grown in the presence of either AR-C155858 or EIPA, but not UK-5099, inhibitor). Each inhibitor remained in the growth medium with half of its initial respective concentration(s) for 24 h more, post-pre-incubation. Survival rates of each cocktail (3-BrPA plus inhibitor) were normalized according to the respective values of inhibitor only. Stock solutions of all three inhibitors (UK-5099, AR-C155858 and EIPA) ( c - e ) were prepared in DMSO. Pre-incubation (for 1.5 h) of T24 with 100 μM EIPA proved significantly detrimental for the cells (data not shown). ( c - e ) Results are reported as mean ± standard deviation of triplicates of three independent experiments. * P < 0.001

Article Snippet: Rabbit polyclonal antibodies against SMCT1 and MPC1 were purchased from Novus Biologicals LLC (Connecticut, USA).

Techniques: Western Blot, Immunofluorescence, Expressing, Incubation, Concentration Assay, Standard Deviation