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. Author manuscript; available in PMC: 2013 Aug 1.
Published in final edited form as: Peptides. 2012 May 22;36(2):151–156. doi: 10.1016/j.peptides.2012.05.006

Fig. 2. Fluorescent staining for the determination of bacteria killed by AG.

Fig. 2

In order to monitor the real-time actions of ghrelin’s bactericidal effects against E. coli, AG in 10 mM NaPi (pH 6.8) solution was incubated (0, 100, and 200 μg/ml AG in 108 cells/ml bacteria suspensions) for 2 hours at 37°C. Immediately after incubation, live and dead bacteria were distinguished by a fluorescent-based staining system (LIVE/DEAD® BacLight™). More specifically, SYTO9 and propidium iodide (PI) were used to stain live cells green and dead cells red, respectively. (A) Representative fluorescent microscopic images of bacteria incubated with or without AG (0 or 200 μg/ml AG in 108 cells/ml bacteria suspensions) are shown. (B) Histogram showing the percentage of dead bacteria stained red with PI. The data (percent of dead bacteria after treatment with 0, 100, and 200 μg/ml AG in 108 cells/ml bacteria suspensions) are shown as the mean ± standard deviation of three independent experiments. Significantly different from control, nontreated bacterial cells (Student’s t-test: * P < 0.05, ** P < 0.01).