Transcription-coupled DNA repair underlies variation in persister awakening and the emergence of resistance

Assessing persister awakening dynamics following antibiotic treatment in E. coli

“…Hence, for populations with lower persister fractions, agarose pads are the preferred solution. Of note, in our set-up with 1% of persister cells, we were able to detect a variation in the awakening lag of the persister cells from 75 (minimal) to 664 min (maximal) within one sample ( Wilmaerts et al., 2022 ).…”

“…Following ofloxacin treatment, strong filamentation is observed in both sensitive cells and persister cells during recovery, resulting from the induction of the SOS response due to double-strand break formation ( Goormaghtigh and Van Melderen, 2019 ; Murawski and Brynildsen, 2021 ; Wilmaerts et al., 2022 ). Some of these filamented sensitive cells will even undergo cell division.…”

“…Strong filamentation is observed following fluoroquinolone treatment, resulting from the induction of the SOS response due to double-strand break formation ( Goormaghtigh and Van Melderen, 2019 ; Murawski and Brynildsen, 2021 ; Wilmaerts et al., 2022 ). After ofloxacin treatment (5 μg/mL) of stationary phase BW25113 cells, filamentation of both sensitive and persister cells can be expected ( Figure 4 ).…”

“…Although the setup is flexible, time-lapse microscopy requires a minimal number of persisters being present. For complete details on the use and execution of this protocol, please refer to Wilmaerts et al. (2022) .…”

“…For complete details on the use and execution of this protocol, please refer to Wilmaerts et al. (2022) .…”