ATP-induced autophagy is associated with rapid killing of intracellular mycobacteria within human monocytes/macrophages-0

-II (16 kDa) by Western blot. Lane 2 represents untreated control THP-1 cells. B. ATP (3 mM) treated MDMs also express LC3-II (lane 2), which is absent in non-treated control cells (lane 1). C. THP1 cells treated with 3 mM ATP for 30 minutes cultured in calcium-free medium (lane 1) and calcium-replete medium (lane 2). LC3-II expression was observed in ATP-treated cells cultured in calcium-replete medium (lane 2) but was absent in cells cultured in calcium-free media (lane 1). D. THP1 cells were treated with different P2 agonists and antagonists. Lane 1 represents untreated control cells, while lanes 2 and 3 represent cells pre-treated with oATP (0.3 mM) and anti-P2Xantibody (3 μg/ml) for 2 hours and 1 hour respectively, prior to treatment with ATP (3 mM for 30 minutes). Lanes 4, 5 and 6 represent cells treated with ATP (3 mM), UTP (3 mM) and bzATP (3 mM) for 30 minutes. The presence of an LC3-II band, indicating cell autophagy, was observed only in lanes 4 and 6.<p><b>Copyright information:</b></p><p>Taken from "ATP-induced autophagy is associated with rapid killing of intracellular mycobacteria within human monocytes/macrophages"</p><p>http://www.biomedcentral.com/1471-2172/9/35</p><p>BMC Immunology 2008;9():35-35.</p><p>Published online 15 Jul 2008</p><p>PMCID:PMC2483257.</p><p></p

ATP-induced autophagy is associated with rapid killing of intracellular mycobacteria within human monocytes/macrophages-3

or without pre-treatment with wortmannin (100 nM/1 hour) or oATP (0.3 mM/2 hours). The kinetics of BCG viability was determined by H-uridine incorporation, monitored at various times post treatment. The figure illustrates the pattern of results obtained from three separate experiments performed in triplicate. The symbols indicate mean values, and vertical bars indicate the standard error. The significance of the various treatment relative to the untreated control are illustrated by: ** = P < 0.005 * = P < 0.05. The results from one H-uridine uptake assay are tabulated as mean counts per minute (CPM) data +/- SEM at each time point examined.<p><b>Copyright information:</b></p><p>Taken from "ATP-induced autophagy is associated with rapid killing of intracellular mycobacteria within human monocytes/macrophages"</p><p>http://www.biomedcentral.com/1471-2172/9/35</p><p>BMC Immunology 2008;9():35-35.</p><p>Published online 15 Jul 2008</p><p>PMCID:PMC2483257.</p><p></p

ATP-induced autophagy is associated with rapid killing of intracellular mycobacteria within human monocytes/macrophages-4

-II (16 kDa) by Western blot. Lane 2 represents untreated control THP-1 cells. B. ATP (3 mM) treated MDMs also express LC3-II (lane 2), which is absent in non-treated control cells (lane 1). C. THP1 cells treated with 3 mM ATP for 30 minutes cultured in calcium-free medium (lane 1) and calcium-replete medium (lane 2). LC3-II expression was observed in ATP-treated cells cultured in calcium-replete medium (lane 2) but was absent in cells cultured in calcium-free media (lane 1). D. THP1 cells were treated with different P2 agonists and antagonists. Lane 1 represents untreated control cells, while lanes 2 and 3 represent cells pre-treated with oATP (0.3 mM) and anti-P2Xantibody (3 μg/ml) for 2 hours and 1 hour respectively, prior to treatment with ATP (3 mM for 30 minutes). Lanes 4, 5 and 6 represent cells treated with ATP (3 mM), UTP (3 mM) and bzATP (3 mM) for 30 minutes. The presence of an LC3-II band, indicating cell autophagy, was observed only in lanes 4 and 6.<p><b>Copyright information:</b></p><p>Taken from "ATP-induced autophagy is associated with rapid killing of intracellular mycobacteria within human monocytes/macrophages"</p><p>http://www.biomedcentral.com/1471-2172/9/35</p><p>BMC Immunology 2008;9():35-35.</p><p>Published online 15 Jul 2008</p><p>PMCID:PMC2483257.</p><p></p

ATP-induced autophagy is associated with rapid killing of intracellular mycobacteria within human monocytes/macrophages-5

Ages of live MDMs infected with GFP-BCG (green) and pre-pulsed with Lysotracker (red) to stain acidic lysosomes. Cells were pre-incubated with anti-P2X7 (3 μg/ml) or wortmannin (100 nM) for 1 hr and then treated with 3 mM ATP for 30 minutes. Note scale bars = 10 um. C. Graph showing the percentage of Lysotracker positive, GFP-BCG containing phagosomes. Histogram shows means ± s.e.m. (n = 25–60 phagosomes). *** = P < 0.01.<p><b>Copyright information:</b></p><p>Taken from "ATP-induced autophagy is associated with rapid killing of intracellular mycobacteria within human monocytes/macrophages"</p><p>http://www.biomedcentral.com/1471-2172/9/35</p><p>BMC Immunology 2008;9():35-35.</p><p>Published online 15 Jul 2008</p><p>PMCID:PMC2483257.</p><p></p

ATP-induced autophagy is associated with rapid killing of intracellular mycobacteria within human monocytes/macrophages-2

Mes (BP). B. Exposure to ATP is associated with the appearance of numerous double-membrane autophagic vacuoles (AV) within the cell cytoplasm. C. A BCG-infected human macrophage (MDM) following treatment with 3 mM ATP for 30 minutes, the bacteria localize within autophagic vacuoles (AV) with clearly demonstrable inner (double black arrows) and outer (double white arrows) membranes. D. A higher magnification image of a number of mycobacteria contained within the inner and outer membranes of an autophagic vacuole.<p><b>Copyright information:</b></p><p>Taken from "ATP-induced autophagy is associated with rapid killing of intracellular mycobacteria within human monocytes/macrophages"</p><p>http://www.biomedcentral.com/1471-2172/9/35</p><p>BMC Immunology 2008;9():35-35.</p><p>Published online 15 Jul 2008</p><p>PMCID:PMC2483257.</p><p></p