Transgenic CL2006 worms synchronized to the L4 stage (early adult stage) were inoculated on NGM plates with or without drugs, and N2 was used as a negative control for Aβ deposition (32 (link)). Following incubation at 20˚C for 48 h, the worms were collected with M9 buffer and fixed in 4% paraformaldehyde/PBS (pH 7.4) (cat. no. BL539A; Biosharp Life Sciences) at 4˚C for 24 h. The worms were then treated with 5% β-mercaptoethanol (cat. no. M828395; Macklin, Inc.), 1% Triton X-100 (cat. no. MB2486; Dalian Meilun Biology Technology Co., Ltd.) and 125 mM Tris (pH 7.4) (cat. no. T8060; Beijing Solarbio Science & Technology Co., Ltd.) at 37˚C for 24 h. The samples were stained with 0.125% thioflavin S (cat. no. S19293; Shanghai Yuanye Bio-Technology Co., Ltd.) in 50% ethanol at room temperature for 2 min, and then with 50% ethanol. The samples were thereafter rinsed and transferred to a glass slide for observation under a laser scanning confocal microscope (LSM900; Carl Zeiss AG). The number of thioflavin S-reactive deposits in the area anterior to the pharyngeal bulb was counted.