Bacterial Growth Assay on NGM Plates

Bacterial growth assays were performed following the method described previously [13 (link)]. OP50 was plated onto solid LB medium and cultured inverted at 37°C overnight. In the next day, a colony was picked and inoculated into liquid LB medium, followed by culturing at 37°C for 16 h. The bacterial culture was diluted in LB medium to OD₆₀₀ = 0.4, and 30 μL of the dilution was added onto NGM plates at 20°C. The plates were rinsed with M9 buffer at 6 different time points (12 h, 24 h, 36 h, 48 h, 60 h, and 72 h) after incubation, and the OD₆₀₀ was measured on a Hitachi U-2910 spectrometer. The experiments were repeated three times independently, and the statistical significance of the growth inhibition was assessed by multiple t-tests.