Fig. 4

Effects of PpSP32 on human THP-1-derived macrophages. A THP-1-derived macrophage viability assessed using the MTT assay was evaluated by incubating cells in the presence or absence of different concentrations of PpSP32 for 24 h and 72 h. Results of three independent experiments are expressed as mean of percentages ± SD of viability according to the control condition. B THP-1-derived macrophages were pretreated with PpSP32 (0.5, 2, or 5 μg/ml) for 48 h, then incubated with 100 ng LPS for 3 h. Whole-cell lysates (30 µg/lane) were then separated, transferred, and incubated with anti-p-IκB-α antibody or anti-β actin. Immunoblots were determined by enhanced chemiluminescence after adding the HRP-conjugated secondary antibodies. Quantification of phospho-IκB level was performed by Image J (version 1.8.0). Data are presented as means ± SD. The P-value was determined by the ANOVA test with multiple comparisons. *Indicates P < 0.05. LPS, lipopolysaccharide; p-IκB-α, phospho I kappa B alpha; Uns, unstimulated; µg, microgram; ml, milliliter; h, hour; PpSP32, Phlebotomus papatasi salivary protein 32