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Fig. 1 | BMC Microbiology

Fig. 1

From: Identification of Mycobacterium avium subsp. hominissuis secreted proteins using an in vitro system mimicking the phagosomal environment

Fig. 1

M. avium secreted proteins within THP-1 macrophages. Differentiated THP-1 cells were infected with M. avium gene constructs fused to a beta-lactamase without secretion signal. After 3 days of infection, FRET-based substrate, CCF2-AM, was loaded into the cells and a shift in fluorescence was observed microscopically (ad) and quantitatively (e). a M. avium transformed with pMV261-BlaC(-) negative control. b M. avium transformed with pMV261-BlaC(+) positive control. Since all 10 of the tested secreted protein constructs produced a color change, we randomly picked MAV_4077 (c) and MAV_0516 (d). Bar, 10μm. e Calculated and normalized fluorescence ratios of the 10 tested protein constructs from the same wells where the fluorescent microscopy was conducted. f. Infection rate was calculated by bacterial CFU at 2 h of initial infection divided by the infection inoculum and normalized for small differences in inoculum between the constructs. g CFU of M. avium recovered during 3 days of infection. Bars represent mean values ± standard deviation. Data shown is representative of three biological replicates. Statistical comparisons: *P <0.05 and **P <0.01 when compared to pMV261-BlaC(+)

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