Quantitative real-time RT-PCR (qRT-PCR) analysis
Total RNA extraction from CD27+IgD+B cells utilized RNeasy Mini Kit (Qiagen, Germany). Reverse
transcription was carried out with the RevertAid First Strand cDNA
synthesis kit (Fermentas, USA) as instructed by the manufacturer,
followed by qRT-PCR assessment. GAPDH and β-actin were employed for
normalization, and the 2-ΔΔCT method was applied for
data analysis(11).