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. 2021 Dec 7;12(6):e03137-21. doi: 10.1128/mBio.03137-21

FIG 2.

FIG 2

Characterization of genes required for survival when OXA-23 is overexpressed. (A) Strain ATCC 17978 Tn7::blaOXA-23 with indicated transposon insertions was spotted on LB agar supplemented with 30 μg/ml kanamycin (Control) and 5 mM IPTG to induce OXA overexpression (+IPTG). (B to D) Growth curves of ATCC 17978 Tn7::blaOXA-23 control and ATCC 17978 Tn7::blaOXA-23 transposon insertions in LB with and without 5 mM IPTG. Six independent transformants were picked directly into 1 ml LB plus 30 μg/ml kanamycin and grown overnight at 28°C. Overnight cultures were diluted to an OD600 of 0.05 in 2 ml LB and supplemented with 5 mM IPTG. Cultures were grown for 8 h shaking at 37°C with OD600 monitored every 30 min. (E) Phase-contrast microscopy of ATCC 17978 Tn7::blaOXA-23 control and ATCC 17978 Tn7:: blaOXA-23 transposon insertions. Samples were collected after 3 h growth after IPTG addition as described above. Cell morphology was visualized using ×1,000 magnification on an Olympus BX51 microscope and imaged using a Lumenera Infinity2 camera. Statistical significance determined by paired two-tailed Student’s t test is indicated by asterisks as follows: *, P < 0.05; **, P < 0.01; ***, P < 0.001.