Figure 12
Effect of protein synthesis inhibition on inducible defensin expression. 16HBE human epithelial tracheal cells (5 × 106) were grown in six well plates for 24 hours. The cells were then pre-treated with 2.5 μg of cycloheximide (CHX) for 1 h, and some samples were then exposed to the different morphotypes of A. fumigatus, either for 6 (Figure 8A) or for 18 (Figure 8B) hours. There was no significant difference in viability between control and treated cells as assessed by staining with trypan blue. Furthermore, the yields of total RNA from the samples were compared and showed no difference. Total RNA was extracted and analysed by RT-PCR. The sizes of amplified products are indicated and were as predicted. GAPDH was uniformly expressed. Complete inhibition of hBD2 and hBD9 expression by the cells exposed to A. fumigatus either for 6 or for 18 hours was observed after pre-treatment of the cells with cycloheximide.