kin-20(ox423) suppressor screen

Methods are similar to those described (Hollopeter et al., 2014). Late L4 Caenorhabditis elegans larvae were mutagenized with 0.5 mM N-ethyl-N-nitrosourea (ENU) for 4hrs at 22C. Animals were washed with M9 buffer, then ~1000 L4 to young adults were pipetted onto a dense lawn of NA22 E. coli grown on 10-cm nematode growth medium (NGM) enriched peptone agar plates, on 66 plates in all. Animals were grown to starvation, then a ~2.5 × 2.5 cm piece of agar was cut from each plate and transferred to new NA22 bacteria on NGM enriched agar plates. In this screen, multiple independent mutagenized populations were generated, each containing enough genetic diversity to give rise to at least one suppressor. The populations were propagated for ~ ten generations, and then one mobile animal was selected from each population.