Fig. 3

Effects of linalool, LEOs prepared at the beginning and at the end of the flowering period and NFκB inhibitors on mRNA (A,C,E,G) and protein (B,D,F,H) levels of pro-inflammatory cytokines IL-6, IL-8, IL-1β and TNFα after P. aeruginosa LPS pretreatment. THP-1 cells were pretreated with 100 ng/mL LPS for 24 h then 500-fold diluted linalool and LEOs or 5 μM luteolin or ACHP were added to the pretreated THP-1 cells for an additional 24 h. DMSO treated cells were used as control of the treated cells. Real Time PCR was carried out with SYBR green protocol using pro-inflammatory cytokine specific primers. β-actin was used as housekeeping gene for the normalization and relative expression of controls was regarded as 1. Pro-inflammatory cytokine secretions were determined using specific ELISA kits according to the manufacturer’s protocols. The columns represent mean values and error bars represent standard deviation (SD) for three independent determinations (n = 3). Real Time PCR and ELISA measurements were carried out in triplicate in each independent experiment. Asterisks indicate p < 0.05 compared to the control. Crosses show p < 0.05 compared to the LPS treatment. Double cross represent statistical significance (p < 0.05) compared to luteolin treatment. Number sign marks p < 0.05 compared to ACHP treatment. The diagrams were created using the same logical structure and the different treatments appeared in the same sequence in each figure in order to improve traceability of the results