Anaplasma phagocytophilum infection modulates expression of megakaryocyte cell cycle genes through phosphatidylinositol-3-kinase signaling
Fig 5
A. phagocytophilum infection increases Akt and NF-κB activation in MEG-01 cells.
(A) LDH assay results performed with cell culture supernatants collected from A. phagocytophilum-infected mock-treated (mock) or A. phagocytophilum-infected LY294002 (PI3K inhibitor)-treated at different days (1, 3, 5, 7) p.i is shown. Values on the Y-axis show the absorbance for LDH release in the cell culture supernatants. The results are from three independent experiments performed in triplicates. All cultures were started simultaneously at the same time. The culture medium was not changed daily. The P value (P<0.05) calculated from Student’s t test was considered significant. (B) Immunoblotting analysis showing levels of phosphorylated Akt and IκB alpha in uninfected (UI) or A. phagocytophilum-infected (I) MEG-01 cells at days 1 and 7 p.i. The levels of actin (detected by immunoblotting) and Ponceau stained gel image for total protein serves as loading control. Densitometry analysis showing levels of Akt (C and D) or IκB alpha (E and F) at days 1 (C and E) and 7 (D and F) p.i., relative to the respective actin bands seen in B.