ESX-1-induced apoptosis is involved in cell-to-cell spread of Mycobacterium tuberculosis
Version of Record online: 2 AUG 2013
© 2013 John Wiley & Sons Ltd
Volume 15, Issue 12, pages 1994–2005, December 2013
How to Cite
Aguilo, J. I., Alonso, H., Uranga, S., Marinova, D., Arbués, A., de Martino, A., Anel, A., Monzon, M., Badiola, J., Pardo, J., Brosch, R. and Martin, C. (2013), ESX-1-induced apoptosis is involved in cell-to-cell spread of Mycobacterium tuberculosis. Cellular Microbiology, 15: 1994–2005. doi: 10.1111/cmi.12169
- Issue online: 15 NOV 2013
- Version of Record online: 2 AUG 2013
- Accepted manuscript online: 15 JUL 2013 03:45AM EST
- Manuscript Accepted: 5 JUL 2013
- Manuscript Revised: 4 JUL 2013
- Manuscript Received: 21 MAY 2013
- Spanish Ministry of Science and Innovation. Grant Number: BES-2006-11950
- Aragón I+D (ARAID)
- Spanish Ministry of Economy and Competitiveness. Grant Numbers: BIO2011-23555, SAF2011-25390
- FP7 European. Grant Numbers: NEWTBVAC 241745, TB-VIR 200973
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.
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.
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.
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.
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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