Regulation of CXCL13 gene expression by NF-κB2 protein. (A-B) Indicated siRNA or pcDNA3-HA-NFKB2 (WT or mutants) were transfected into total PBMCs from healthy donor control. Next day, cells were washed and stimulated with Dynabeads-CD3/28 for 48 hours. RNA was prepared and CXCL13 gene expression was analyzed by real-time PCR. A fold change was calculated for NFKB2 siRNA or mutants with control vector (A) or WT (B) transfected cells, normalize to 1. CXCL13 was not detected in unstimulated samples. Decreased NF-κB2 (A) or overexpressed WT or mutant protein expression (B) was confirmed by western blot. Results shown are means + standard error of the mean of 3 independent experiments. (C) CD4 T cells were enriched using stem cell negative selection kit and cells were stimulated with Dynabeads-CD3/28 for 48 hours. Levels of CXCL13 mRNA in activated CD4 T cells from indicated patients were measured by real-time PCR using the probe for CXCL13 and normalized to 18S rRNA. CXCL13 expression was not detected in A1 due to low cell number. Data are means value of replicates from indicated patients. For relative gene expression, all data were normalized to the paired normal control.