Figure 3

FOXJ1-mediated expression of cilia-related genes and enhancement of their promoters in primary human basal cells transfected with either control EGFP or FOXJ1 expression plasmids. All data is based on TaqMan quantitative real-time RT-PCR 48 hr after transfection and a gene was deemed “undetectable” if not detected after 40 cycles of assessed undiluted cDNA. A. Basal cell markers KRT5, KRT14 and TP63. B. Secretory cell markers MUC5AC, MUC5B and SCGB1A1. C. Ciliated cell-associated genes, including centrin 2 (CETN2); dynein, axonemal, heavy chain 11 (DNAH11), dynein, axonemal, intermediate chain 1 (DNAI1); dynein, axonemal, light intermediate chain 1 (DNALI1); EF-hand domain, C-terminal, containing 1 (EFHC1); sperm associated antigen 6 (SPAG6), tektin 1 (TEKT1), TEKT2 and tubulin, alpha 1a (TUBA1A). D. Shown is firefly luciferase activity in basal cells transfected with control EGFP or FOXJ1 expression plasmids together with firefly luciferase reporter gene plasmids driven by promoters of the ciliated cell-associated genes DNALI1, DNAI1, SPAG6 and TEKT1. Controls include the promoter for the basal cell marker KRT14 and a random sequence. The data were normalized for transfection efficiency per well by Renilla luciferase activity from co-transfected Renilla luciferase plasmid (pTK-RL). Bars represent mean ± standard error of pooled data from replicates of a minimum of three individual experiments with cells from three different subjects.