Figure 6.

Replication of IAV subtypes and inflammatory responses of A549 cells. (A) Following entry inhibition, replication of IAV subtypes (H1N1 and H3N2) is affected by factor H and VCP in human A549 cells. M1 expression was monitored in A549 cells challenged with either H1N1 or H3N2 IAV subtypes (MOI 1) at 6 h. A549 cells were challenged with H1N1 and H3N2 pre-incubated with or without factor H and VCP (40 μg/ml). Cell pellets were harvested at 6 h post-infection. 18S rRNA was used as an endogenous control. Significance was determined using the unpaired one-way ANOVA test (**p < 0.01, and ***p < 0.001) (n = 3). (B) Transcription expression profile of cytokines and chemokines produced by A549 in response to H1N1 and H3N2 challenge with and without factor H. 18S rRNA expression was used as an endogenous control. Cells only was used as a calibrator sample to calculate relative quantitation (RQ); RQ = 2^−ΔΔCt. Experiments were conducted in triplicates, and error bars represents ± SEM. Unpaired one-way ANOVA test was used calculate the significance (*p < 0.05, **p < 0.01, and ***p < 0.001) (n = 3). (UT, untreated sample; T, treated sample). (C) Expression levels of type I interferon alpha (IFN-α) following 6 h treatments with factor H and VCP. 18S rRNA was used as an endogenous control. RQ = 2^−ΔΔCt was used to calculate the RQ value. Significance was determined using the unpaired one-way ANOVA test (**p < 0.01) (n = 3).