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After washing another time, the cells were dehydrated in a graded series of alcohol (30%, 50%, 70%, 80%, 90%, 95%, and 100%) before being embedded in EMBed 812 (SPI, United States), and ultrathin sections (6080 nm) were prepared using a Leica ultramicrotome (Leica Microsystems, United States) and fished out onto the 150-mesh cuprum grids with a formvar film

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For experiments, the synchronized L1 larvae were placed on the new NGM plates containing OP50 and incubated at 20C and allowed to grow to L3 larvae