Quantitative real-time PCR
Total RNA from B cells was isolated by the Takara PrimeScript RT reagent
kit according to the manufacturer’s protocol. Quantitative real-time PCR
(qRT-PCR) was performed using the Bio-Rad CFX96 system. Each sample was
processed and analysed in duplicate. The primers used for qRT-PCR are
listed in supplemental Table S2. To quantify the relative expression of
each gene, Ct values were normalized to the endogenous reference values
(ΔCt = Ct target-Ct GAPDH) and compared with a calibrator using the ΔΔCt
method (ΔΔCt = ΔCt sample-ΔCt calibrator).