Fig. 4.
The effect of long-term pre-culturing on persister resuscitation. a-b Cells from 9-day overnight pre-cultures were inoculated in fresh media, cultured until OD600 = 0.25, and then treated with ampicillin for 3 h or 16 h. After ampicillin treatments, cells were transferred to LB broth to monitor persister resuscitation with a flow-cytometry (N = 4). c Cells during the antibiotic treatments were plated to obtain the bi-phasic kill curve (N = 4). d The doubling time of the resuscitating cells after 3-h ampicillin treatments (N = 4). e The doubling time of the resuscitating cells after 16-h ampicillin treatments (N = 4). f-g Comparison of non-lysed cell levels in short-term (1 day) and long-term (9 days) pre-cultures (N = 4). * indicates a significant difference between short-term and long-term pre-cultures (P < 0.05). h-i Resuscitating cell levels after 3- and 16-h antibiotic treatments (N = 4). * indicates a significant difference between flow-cytometry (to = 0) and agar plate cell counts (P < 0.05). # indicates a significant difference between flow-cytometry (to = 60 min) and agar plate cell counts (P < 0.05). Error bars represent the standard deviations. ONC: Overnight pre-culture