PU.1 overexpression increases NLRC4 gene expression in relation to the rs385076 C allele. HEK293A cells were transfected with pGL4.10[luc2] plasmid, containing either rs385076 C (shown in red) or T allele (shown in green) and the putative PU.1 binding site, as well as the pVITRO2-PU.1 plasmid. Expression of yellow fluorescent protein after transfection of an empty pEYFP-N1 plasmid was used to control for transfection efficiency. PU.1 overexpression significantly increased luciferase activity of the rs385076 C allele but not the T allele in the plasmids. Without PU.1 overexpression, there was no significant difference between C and T alleles. Cells with 0.125 μg/mL (4.24-fold, P=3×10−4), 0.25 μg/mL (3.34-fold, P=3×10−4) and 0.5 μg/mL (3.5-fold, P=5×10−4) of the pVITRO2-PU.1 plasmid showed significant increases of luciferase activity for the C allele compared to the T allele. ns = not significant, *** P <0.001; n=4.