Fig. 5: AVAN enhances FOXO3a expression in nucleus.

A, B FOXO3a expression during virus infection in A549 cells were measured by qRT-PCR. A549 cells were transfected with AVAN plasmids (A) or siRNA (B) (n = 3; means ± SEM; *p < 0.05; **p < 0.01; ***p < 0.001). C Enrichment of AVAN in ChIRP assay analyzed by qRT-PCR, U1 as a negative control. ChIRP assay showing that AVAN binds directly to the FOXO3a promoter DNA in BJ501-uninfected (D) and -infected (E) A549 cells. P1–P10 represent the different regions in FOXO3a promoter or gene body locus. (F–K) H3K4me3 and H3K27ac levels and Pol II binding of the FOXO3a promoter were analyzed through ChIP followed by qRT-PCR in BJ501-infected A549 cells. Endogenous RIP of SET1A (L), CBP (M), and Pol II (N) in BJ501-infected cells using specific or anti-IgG antibodies. The relative enrichment fold of AVAN was calculated by qRT-PCR (n = 3; means ± SEM; *p < 0.05; **p < 0.01). O, P Transwell assay of neutrophil migration in FOXO3a-overexpressing or FOXO3a-knockdown A549 culture supernatant (n = 3; means ± SEM; *p < 0.05; **p < 0.01). (Q, R) IL-8 expression in FOXO3a-overexpressing (N) or FOXO3a-knockdown (O) A549 cells were measured by qRT-PCR (n = 3; means ± SEM; *p < 0.05; **p < 0.01). S IL-8 expression in AVAN-overexpressing or/and FOXO3a-knockdown A549 cells were measured by qRT-PCR (n = 3; means ± SEM; **p < 0.01; ***p < 0.001).