IL-1α mRNA and protein expression in HeLa cells is regulated by seminal plasma via the EP2 receptor, EGFR and PI3K pathways. IL-1α mRNA (A and B) and protein (C) as determined by qPCR and ELISA analysis, respectively. HeLa cells were treated for 4 (A and B) and 16 hours (C) with seminal plasma (1:50) or vehicle in the absence/presence of EP2 receptor antagonist [AH-6809; 20 μM] and chemical inhibitors to EGFR kinase [AG-1478; 100 nM], PI3 kinase [LY-294002; 25 μM], PTGS1 [SC-560; 15 μM] or PTGS2 [NS-398; 8 μM]. Data are represented as mean ± SEM from 5 independent experiments. Paired T-tests were conducted on the untransformed means of the replicates between SP and control and unpaired T-tests performed on SP versus SP and inhibitor after conversion to fold increases. *, **, and *** indicates significance at P < 0.05, P < 0.01, and P < 0.001, respectively.