Fig. 3.

BAFF expression was inhibited by the treatment with KR33426. (A-D) MH7A cells were transfected with 0.25 µg (A) or 0.5 µg (A-D) pGL3-BAFF-Luc DNA. Luciferase activity of different DNA concentration of BAFF promoter (A) or each time point (B) was measured by using luminometer. Cells were incubated by the stimulation with TNF-α for various time points and luciferase activity was measured (C). Cells were stimulated with TNF-α in the presence or absence of KR33426 and luciferase activity was measured (D). (E-G) MH7A cells were stimulated with TNF-α in the presence or absence of KR33426 for 6 h. RNA was isolated with Trizol. BAFF transcripts were measured by RT-PCR (E) and realtime qPCR normalized to β-actin expression (F). Cell lysates were prepared and western blotting was used to detect BAFF (G). Data were the representative of four experiments (A-G). Data in the bar graph represent the means ± SED. **p<0.01, significant difference as compared to TNF-α-untreated control. &p<0.05, &&p<0.01, significant difference from TNF-α-treated and KR33426-untreated control (D, F).