Transcriptional Response of Lung Epithelial Cells to Aspergillus Proteins

Mouse lung epithelial cells (MLE12) were obtained from ATCC (Manassas, VA, USA); 3 × 10⁵ MLE12 cells were plated in 24-well plates and incubated overnight at 37°C. The cells were stimulated with rAc-PF and Aspergillus proteins for 3 h. Next, MLE12 cells were collected with 1 mL of QIAzol (Qiagen, Hilden, Germany), and RNA extraction was conducted in accordance with the manufacturer’s protocols for transcription of 2 μg of RNA. Real-time PCR was performed to determine the levels of macrophage-derived chemokine (MDC; CCL22), eotaxin (CCL11), thymic stromal lymphopoietin (TSLP), and IL-25 RNA. GAPDH was used as an internal reference. The primers and PCR conditions used have been described previously [25 (link), 26 (link)].