Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Impact of keto leucine and isoleucine on CHO cell central carbon metabolism and performance in fed-batch and steady-state perfusion

doi: 10.3389/fbioe.2026.1708088

Figure Lengend Snippet: Abundance profiles of BCAA-derived metabolites, which were identified in spent media of simulated perfusion processes with CHOK1 GS and CHO DG44 (mean ± SD, n = 2). (A) α-Hydroxy acid of leucine (HL) and α-hydroxy acid of isoleucine (HI). Due to the close elution of the isomers, this feature is computed as a sum parameter of HI and HL. When the levels of HL strongly exceeded the HI levels, the peaks could not be integrated separately. (B) HI (when separation from HL peak was possible). (C) HV, 2-hydroxy-3-methylbutyric acid. (D) dcKI/dcKL, 3-methylbutanoic acid and 2-methylbutanoic acid, the decarboxylation products of keto leucine and keto isoleucine, respectively. Due to co-elution, this feature was determined as a sum parameter. (E) HMB, 3-hydroxyisovaleric acid, a by-product of Leu metabolism. (F) HMH, (3S)-3-hydroxy-5-methylhexanoic acid.

Article Snippet: 1,2- 13 C 2 - and U- 13 C 6 -D-glucose, d 7 -2-hydroxy-3-methylbutyric acid, d 8 -3-methyl-2-oxopentanoic acid, d 3 -4-methyl-2-oxopentanoic acid, and d 3 -L-2-hydroxy-4-methylpentanoic acid were sourced from Toronto Research Chemicals (Toronto, ON, Canada). d 10 -2-Hydroxy-3-methylpentanoic acid (mixture of all four diastereomers) was synthesized internally from d 10 -isoleucine sourced from CDN Isotopes (Teddington, United Kingdom).

Techniques: Derivative Assay, Co-Elution Assay

Journal: Journal of Neuroinflammation

Article Title: SYK regulates Schwann cell metabolic reprogramming to promote axonal regeneration in immune-mediated neuropathy

doi: 10.1186/s12974-025-03624-y

Figure Lengend Snippet: SYK deficiency impairs glycolytic flux in SCs. ( A ). ECAR profiles measured by Seahorse experiments. ( B ). Quantification of basal glycolysis, glycolytic capacity, and glycolytic reserve in ECAR. n = 4 independent biological experiments. ( C ). Glucose uptake measured with 2-NBDG by flow cytometry. n = 4 independent biological experiments. ( D ). Intracellular levels of glucose-6-phosphate (G6P), pyruvate and lactate were measured by chemiluminescence. n = 4 independent biological experiments. ( E ). Protein levels of GLUT1, GLUT4, HK2, PFK-1, PFKFB3, PKM2 and LDHA in SCs were measured by western blot. n = 3 independent biological experiments. ( F ). Immunofluorescence staining and quantification of GLUT1, HK2, PFK-1, and PFKFB3 in SCs. Scale bar, 20 μm. N = 4 independent biological experiments, with n ≥ 6 fields/experiment. ( G ). Schematic depicting SYK deficiency-induced downregulation (red arrows) of glycolytic transporters, enzymes, and metabolites in SCs

Article Snippet: SCs were incubated with 100 μM 2-[N-(7-nitrobenz-2-oxa-1,3- diazol-4-yl)amino]−2-deoxy-D-glucose (2-NBDG) (MCE, Cat#HY-116215) for 30 min at 37 °C in the dark.

Techniques: Flow Cytometry, Western Blot, Immunofluorescence, Staining