Fig. 6.

IRF1 Knockdown inhibited HNP1 promotion of expression of proinflammatory cytokines in CAL-1 cells. CAL-1 cells transfected with control (Ctl) or IRF1 (IRF1 KD) siRNAs were cultured at 37 °C overnight. Subsequently, a portion of the cells was used for the preparation of whole cell lysate, while the rest of the cells were treated with HNP1 (final concentration = 2 μM) and/or ODN (final concentration = 1 μM) for 5 h at 37 °C before extraction of total RNAs. A, Identical amounts of Ctl and IRF1 lysates were separated on a SDS-PAGE gel, transferred onto a PVDF membrane, and probed by Western blot using anti-IRF1 antibody (upper panel). The same membrane was subsequently stripped and re-probed with anti-β-actin antibody (lower panel). B-D, qPCR quantitation of the mRNA levels of IFNα (B), IFNβ1 (C), and IL-6 (D) in Ctl and IRF1 KD CAL-1 cells. Shown is the average (mean ± SD) of three independent experiments. *p < 0.05 by Student’s t test.