Extended Data Fig. 9: Ldhafl/flAicdaCre/+ mice can mount a robust GC response.
From: Distinct metabolic requirements regulate B cell activation and germinal center responses
Ldhafl/flAicdaCre/+ and Ldha+/+AicdaCre/+ (control) mice were immunized intraperitoneally with NP-CGG, boosted at d10, and analyzed at d14. a, Viability of GC B cells as measured by cells negative for Zombie Red viability dye. b, Cell size of viable GC B cells as measured by quantifying the MFI of forward scatter (FSA). c, Mitochondrial mass was quantified by measuring the MFI of MitoTracker™ Green FM Dye among the GC and non-GC B cells. d, Frequency of dysfunctional mitochondria was measured by quantifying the fraction of live GC and non-GC B cells negative for MitoTracker™ Red CMXRos. e, PCR of genomic DNA of GC and non-GC B cells sorted from spleens of mice of the indicated genotypes. f, Immunoblotting of whole cell extracts prepared from GC and non-GC B cells sorted from the Peyer’s patches and spleens of mice of the indicated genotypes to detect LDHA and α-tubulin (control) proteins. g, Expression of Ldha and Ldhb in sorted GC B cells. Bars represent mean. Each datapoint represents a single mouse (n = 3). The box plot depicts the first quartile, third quartile, and the median of the dataset. Values that fall within 1.5 times the interquartile range above the third quartile and below the first quartile are represented by the whiskers. The gating strategy for the sorting of GC B cells is the same as that in Extended Data Fig. 6a. For panels a and b, n = 8 for each genotype; data is representative of two independent experiments. For panels c and d, n = 5 for each genotype and data is representative of two independent experiments. For panels e, and f, data is representative of 2 independent experiments with 3 to 5 mice per experiment.
