Analysis of mRNA levels for HBD2 and HBD9 in HNT primary culture cells exposed to A. fumigatus organisms. Primary epithelial HNT cells (5 × 106) were grown in six well plates for 48 hours. The cells were then exposed to the different morphotypes of A. fumigatus or latex beads for 18 h. Cells were cultivated in a control well in the absence of A. fumigatus or the latex beads. Isolation of total RNA and synthesis of cDNA was performed as described in Methods. Specific primer pairs and the conditions of real time PCR are described in Table 2. The level of mRNA for defensins was measured in total RNA preparation by quantitative real time PCR as described in Methods. Expression of all genes was normalised to the expression of the endogenous reference gene GAPDH. The expression value in control cells was used as the baseline. Means followed by the same letter are not significantly different.