Skip to main content
. 2014 Dec 22;14(6):307–320. doi: 10.4110/in.2014.14.6.307

Figure 3.

Figure 3

Mycobacterium scrofulaceum-114R strongly increases the production of proinflammatory cytokines through NF-κB signaling in BMDMs. (A) BMDMs were transduced with NF-κB p65 adenovirus luciferase construct (20 PFU/cells) for 36 h and then infected with M. scrofulaceum-ATCC, -113S, -114R (MOI=1, 5, 10) for 4 h. Cell lysates were harvested and luciferase activity was measured. (B) BMDMs were infected with M. scrofulaceum strains (MOI=5) for 30 min and then immunolabeled with anti-NF-κB p65 antibody and anti-rabbit-AlexaFluor 488 (green), and the nuclei were stained with DAPI (blue). Representative immunofluorescence images (left panel) and the average mean fluorescence intensity of cells exhibiting NF-κB nuclear translocation (right panel) are shown. (C) BMDMs were preincubated in the presence or absence of BAY11-7082 (BAY; 0.3, 1, 3µM) or CAPE (1, 5, 10µM) for 45 min prior to infection with M. scrofulaceum strains (MOI=5). The mRNA expression and production of TNF-α and IL-6 were evaluated by semi-quantitative RT-PCR. The data show the mean±SD from three independent experiments. U; uninfected control, SC; solvent control (0.1% DMSO).