Fig. 4.
Extracellular ATP is packaged inside MVs from of S. vesiculosa M7T. MVs were collected from a 48-h TSB culture at 15 °C and 180 rpm. MVs were split into three equal aliquots: one was treated with 0.1% Triton X-100 to lyse the MVs and measure total MV-associated ATP; the second was treated with the apyrase enzyme (2 μg/ml, 30 min, 30 °C) to deplete available ATP outside the MVs and washed with PBS, and then outer-membrane vesicles were lysed with 0.1% Triton X-100 to measure the ATP inside the MVs; in the third, MVs were first lysed with 0.1% Triton X-100 and then treated with apyrase (2 μg/ml, 30 min, 30 °C) to eliminate total ATP. ATP concentration of all three aliquots was measured by the BacTiter-Glo Microbial Cell Viability Assay. n = 3. *P < 0.05 vs the other treatments