Fig. 3

Worm sorting settings based on gating region and Profiler feature. a COPAS Biosort conditions optimised for the sorting of homozygous phb-2(tm2998);Phsp-6::GFP at the L2 stage. The upper panel reflects the gating region based on the extinction peak height (ExtPH) and the extinction peak width (ExtPW) selecting the worm population. The lower panel shows the worm distribution based on green parameters (green peak height (green PH) and green peak width (green PW)). b Utilising the Profiler II software to distinguish between phb-2(tm2998)/mIn1;Phsp-6::GFP and phb-2(tm2998);Phsp-6::GFP larvae using green parameters. The phb-2(tm2998)/mIn1;Phsp-6::GFP worms show two peaks in the green profile corresponding to the two lobes of the pharynx, have a green PH above 10,000 and are excluded (bottom panel). The phb-2(tm2998);Phsp-6::GFP larvae show a profile without pronounced peaks, with a green PH ranging from 700 to 10,000 and green PW above 120, and are accepted and sorted (top panel). The picture includes a balanced heterozygous phb-2(tm2998)/mIn1;Phsp-6::GFP expressing Pmyo-2::GFP in the pharynx and two phb-2(tm2998);Phsp-6::GFP homozygous deletion mutants with induced Phsp-6::GFP expression imaged at the moment of sorting, approximately 48 h after synchronisation