Extended Data Fig. 3: Cellular homeostasis in LDHA-deficient activated B cells.
From: Distinct metabolic requirements regulate B cell activation and germinal center responses
a, Uptake of the fluorescent glucose analog 2-NBDG. Naïve B cells (n = 5 for each genotype) were activated ex vivo with LPS + IL4 for 48 h, incubated with 11 mM of 2-NBDG for 15 min, and 2-NBDG uptake was quantified by flow cytometry. Data is representative of two independent experiments. b, Splenic B cells from Ldhafl/flCd23Cre (n = 3) and Ldha+/+Cd23Cre (control, n = 3) mice were activated ex vivo as indicated for 48 h and intracellular ATP level was determined. Data is representative of two independent experiments. c, d, Splenic B cells from Ldhafl/flCd23Cre and Ldha+/+Cd23Cre (control) mice were labeled with Cell Trace Violet (CTV) dye, activated ex vivo for 48 h with LPS + IL4, and CTV dilution was determined by flow cytometry as a measure of cell proliferation. Data shown is representative of three independent experiments. d, Splenic B cells (1 × 106) were harvested from mice of the indicated genotypes (n = 7 for each group), stimulated with LPS + IL-4 and cell numbers were quantified at the indicated time points. e, Splenic B cells (1 × 106) harvested from mice of the indicated genotypes (n = 4 for each group) were stimulated with LPS + TGFβ + anti-IgD ex vivo and cell numbers were determined at 96 h post-activation. Data is representative of two independent experiments. Bars represent mean ± s.e.m. *p ≤ 0.05, ****p ≤ 0.0001 by unpaired, two-tailed t-test.
