Magic red cathepsin b kit

The Magic Red Cathepsin B Kit (Immunochemistry Technologies, Bloomington, MN, USA) was used, based on manufacturers’ instructions and previous descriptions of use in C. elegans experiments [40 (link)]. In brief, Magic Red reagent dye dissolved in DMSO and further diluted with water was spread over 3 mL NGM plates previously seeded with OP50 E. coli, giving a final 1X concentration of the dye. Once the liquid had dried, hypochlorite-synchronized L4 larvae or mated day 6 adults (previously mated at a 1:2 ratio with fog-2(q71) males from the L4 stage to day 1 of adulthood) were placed on the plates and incubated at 20°C overnight in th e dark, then cleared of residual dye by being placed on fresh plates seeded with OP50 E. coli for 35 minutes prior to imaging. The Magic Red reagent contains a target sequence peptide that is cleaved by cathepsin B to produce a red fluorescent product; red fluorescence intensity was measured using Nikon Elements Advanced Research software.