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cmi12169-sup-0001-si.tif945K

Fig. S1. Apoptosis induced on THP-1 cells by MT103. THP-1 cells were infected with GFP-expressing MT103 bacteria and stained 72 h post infection with Hoechst 33342. A representative image is shown in the figure.

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Fig. S2. Apoptosis induced on MH-S cells by H37Rv and H37Ra strains. MH-S murine macrophages were mock-treated or infected with 5–10 bacteria (H37Rv or H37Ra strains) per cell. At 72 h post infection, apoptosis was determined by flow cytometry with AnnexinV and 7-AAD staining. Data in the graphs are represented as mean ± SD of two independent experiments.

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Fig. S3. Presence of mycobacteria in spleen from infected mice. Groups of five C57BL/6 mice were intranasally infected with approximately 1000 cfu per mouse of MT103, MTBVAC, BCG or BCG::RD1 strains. At 28 days post infection, animals were humanely sacrificed and spleen harvested for in vivo studies. Colony-forming units were determined. Representative data of two independent studies are shown. Statistical analysis was done with one-way anova and Bonferroni's post-test. *, **, *** = statistically significant; *P < 0.05; **P < 0.01; ***P < 0.001.

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Fig. S4. Ziehl-Neelsen and F4/80 immunochemical staining of mouse infected lungs. Mice were intranasally infected with a low dose (approximately 1000 cfu) of the indicated strains.

A. Mycobacteria presence was evaluated by Ziehl-Neelsen staining. Arrows indicate bacteria localization.

B. Macrophage infiltration was determined by immunochemical staining of F4/80 macrophage-specific marker. Representative images (600× magnification for ZN and 100× for F4/80 staining) of infected lungs are shown.

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Fig. S5. Apoptosis induced on MH-S cells by staurosporine. MH-S murine macrophages were incubated for 20 h with the indicated concentrations of staurosporine. Apoptosis was determined by flow cytometry with AnnexinV and 7-AAD staining. A representative experiment of two independent studies is shown.

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