Our lab uses high-throughput methods and develop systematic approaches to probe how environmental cues and genetic background influence antibiotic activity against Gram-negative bacteria. I will show two pioneer examples; in the first we show how a dedicated phage defence system unexpectedly enhances antibiotic activity of antifolates against pandemic Vibrio cholerae. In the second, we combine an experimental screening approach with taylored data analysis to investigate transcriptional regulation of the major drug transporters in Escherichia coli across ~100 small molecules. Our findings show that several chemically diverse and previously unknown compounds trigger transport remodelling beyond what is currently known. Ultimately, our findings contribute to better understanding of bacterial biology in the context of antibiotics, and therefore support future improved treatment designs.