Fig. 2

Independent validation of transcriptional responses of selected protein targets. HBEC-3KT were stimulated with either IL-17A/F (50 ng/mL), TNF-α (20 ng/mL) or IFN-γ (30 ng/mL), or cytokines combinations as indicated, for 6 h. mRNA was isolated and transcriptional responses evaluated by quantitative real-time PCR for (A) antimicrobial HDP LCN-2 (NGAL2) and Elafin (PI3), (B) neutrophil chemokines GROα (CXCL1) and IL-8 (CXCL8), and (C)MMP13. Fold changes (y-axis) for each gene was normalized to 18S RNA, and calculated compared to unstimulated cells normalized to 1, using the comparative ΔΔCt method. Results are shown as boxplots, wherein bars show median and IQR, and whiskers show minimum and maximum values. Each data point represents an independent experimental replicate (n = 4). Fisher’s LSD test for one-way analysis of variance (ANOVA) was used to determine statistical significance (*p < 0.05, **p < 0.01). The dashed line represents normalized baseline value of 1 for unstimulated cells