porcine brain tubulin  (Cytoskeleton Inc)

Journal: Molecular Metabolism

Article Title: A targeted metabolomic method to detect epigenetically relevant metabolites

doi: 10.1016/j.molmet.2026.102342

Figure Lengend Snippet: Distribution of acetyl-CoA isotopomers following [U–13C]glucose and [U–13C]glutamine incubation with an isogenic gain-of-function mutation (R132H) in the metabolic enzyme Isocitrate Dehydrogenase 1 (IDH1). U-87 MG human brain immortalized cell line, either wild-type or harboring the IDH1-R132H mutation, was incubated with [U– 13 C]glucose or [U– 13 C]glutamine for 20 and 72 h. The x axis represents the different acetyl-CoA isotopomers, showing the number of incorporated 13C atoms, with their origins within the acetyl-CoA structure noted in parentheses (see also ). The y axis indicates the percentage abundance of each isotopomer population, with n = 5 per condition after correction for natural abundance (refer to the Methods section 4.6.3). Statistical significance was assessed using Student’s t-test, with p -values represented as follows: ns: not significant ( p > 0.05); ∗: p ≤ 0.05. Because the M+2 labeling of the acetyl group may arise from either oxidative decarboxylation of pyruvate or reductive carboxylation of citrate, these two indistinguishable sources are jointly annotated as M+2 (Pyr/Cit). Abbreviations: Pyr/Cit, pyruvate- or citrate-derived M+2 (indistinguishable by MRM transitions); Gly, Glycine; For, Formate; R5P, Ribose-5-phosphate. # in x-axis indicates isotopomers with the same number of labeled carbon atoms incorporated, but with different origin and final moiety.

Article Snippet: The U-87 MG human brain immortalized cell line (ATCC® HTB-14TM) (wild type) or the R132H (IDH1 mutant) were plated at a density of 1 million cells per T75 plate in 10 mL complete ATCC Eagle’s Minimum Essential Medium (EMEM) 30-2003 medium, with 5 replicates per condition.

Techniques: Incubation, Mutagenesis, Labeling, Derivative Assay