NTHi whole cell lysate (WCL) up-regulates β-defensin 2 (HBD-2) gene expression in immortalized human mucosal epithelial cells (HMEEC-1). NTHi treatment of HMEEC-1 (A) and A549 epithelial cells (B) increased the mRNA levels of β-defensin 2. The x-axis shows the time during which the cells were exposed to NTHi WCL (5 μg/ml). The y-axis represents the signal derived from quantitative PCR on total RNA extracted from the cells. (C), NTHi also up-regulates mouse β-defensin 2 (mBD-2) mRNA expression in the mouse middle ear, in vivo. NTHi WCL (10 μl) was transtympanically inoculated into the middle ears of anesthetized C57BL/6J mice. Animals were sacrificed at the indicated time points and total RNA extracted from the middle ear mucosa (n = 3). Values are given as mean ± standard deviation.