Serum opsonization of Schu S4 leads to inhibition of NF-κB p65 phosphorylation and translocation into nucleus during phagocytosis by macrophages.

<p>(A) hMDMs were infected with pre-opsonized (10% autologous serum) or non-opsonized Schu S4 at an MOI of 50∶1 in RHH. Infection was synchronized by centrifuging at 250×g for 10 min at 4°C, and incubated at 37°C for different time points (15′, 30′ and 60′). MDM lysates were subjected to Western Blot using Phospho NF-κB p65 or total p65 primary antibodies. (B) Cumulative data of band intensities from n = 2 (* p<0.05). (C) hMDMs were fixed after 1 hr of infection with pre-opsonized or non-opsonized Schu S4 at an MOI of 50∶1 and NF-κB p65 translocation was examined by immunofluorescence microscopy with rabbit NF-κB p65 antibody followed by anti-rabbit AF 594 antibody. Nuclei were stained with DAPI. NF-κB p65 translocation was scored using a total of 300 cells per experiment. The graph shown represents cumulative data from two independent experiments. The data were analyzed by a two-tailed Student t-test (** p<0.005, Student t-test).