Extended Data Fig. 2: LDHA governs aerobic glycolysis in B cells.
From: Distinct metabolic requirements regulate B cell activation and germinal center responses
a, Schematic representation of mito stress assays performed in a Seahorse analyzer. The mito stress assay measures the oxygen flux of cells cultured in glucose-sufficient media supplemented with glutamine and sodium pyruvate. During this assay, the oxygen consumption rate (OCR) is first measured at baseline and after the sequential addition of oligomycin (which inhibits ATP synthase and reduces OCR), FCCP (an uncoupling agent that disrupts the mitochondrial membrane potential without inhibiting the electron transport chain and allows oxygen consumption to reach a maximum), and a mixture of rotenone and antimycin A (AA) (which shuts down mitochondrial respiration). b, Representative graph of OCR of naïve B cells at baseline and following the indicated perturbations. c, Quantification of OCR and of spare respiratory capacity (SRC) of naïve splenic B cells from Ldhafl/flCd23Cre and Ldha+/+Cd23Cre (control) mice. SRC was calculated as the difference of FCCP-stimulated maximum OCR and basal OCR. d, e, Naïve splenic B cells were activated with LPS + IL4, anti-CD40 + IL4 or anti-CD40 + IL4+anti-IgM for 48 h and OCR and SRC was quantified. p values were calculated using unpaired, two-tailed t-tests. For panels b–e, n = 6 for each indicated genotype and representative of three independent experiments. Data points and bars represent mean ± s.e.m.
