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. 2021 Jan 27;11:2393. doi: 10.1038/s41598-021-81557-1

Figure 2.

Figure 2

P. aeruginosa is eliminated by murine primary BMDM inside LAMP1-positive vesicles. (a) Intracellular bacterial survival over time was measured by standard antibiotic protection assays followed by CFU counting. Data presented as mean of CFUs ± SEM were normalized to time 0 (100%). This assay was done in the absence (−AC) or presence of apoptotic cells (+AC). Three independent experiments were performed. No significant differences were found by unpaired t test. (b) Phagocytic Index. Cells were incubated with the targets for 30 min. The index was calculated as the average number of bacteria internalized per cell in the presence or absence of apoptotic cells. Data were calculated as mean ± SEM and normalized to the condition with only bacteria (100%). PAK bacteria alone, PAK + AC PAK with apoptotic cells concurrently. Three independent experiments were performed. No significant differences were found by unpaired t test. (c) P. aeruginosa and apoptotic cells localize to LAMP1-positive vesicles. BMDM were fed for 50 min with PAK-mCherry, CFSE-labeled apoptotic cells, or both concurrently. Confocal projected Z-stacks showing LAMP1-positive vesicles containing bacteria and apoptotic material. Scale bars 10 µm. Green: CFSE-labeled apoptotic cells. Red: PAK-mCherry. Magenta: LAMP1.