L4 homozygote larvae were picked and aged to adults. The mouse α-dpMPK-1 (Sigma) was used as a primary antibody (1∶1000). Mouse α-tubulin (1∶000; DSHB) was used as a loading control. Secondary antibodies used were α-mouse antibody conjugated to horseradish peroxidase (HRP; 1∶10,000). PBST-5% milk was used for incubation and blocking. Quantification was done on FIJI [89] (link).
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