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. 2021 May 21;297(1):100813. doi: 10.1016/j.jbc.2021.100813

Figure 5.

Figure 5

ASM overexpression reduces cholesterol accumulation and increases autophagic flux in NPC1 KO HeLa cells.A, ASM activity in HeLa NPC1 KO cells transfected with empty vector (myc-tag) or SMPD1-myc at 24 h post transfection. B, representative confocal images and filipin intensity quantification in empty vector or SMPD1-transfected HeLa NPC1 KO cells at 24 h post transfection. N ≥ 70 cells/condition. C, representative epifluorescent images and quantification of number of LC3 puncta in NPC1 KO cells expressing vector CTR or SMPD1 at 24 h post transfection. N ≥ 55 cells/condition. D, representative epifluorescent images and quantification of p62 puncta in NPC1 KO cells expressing vector CTR or SMPD1 at 24 h post transfection. N ≥ 35 cells/condition. All graphed data show mean ± SD. ∗∗∗p < 0.001 versus vector control in two-tailed Student’s test. In all images, arrows indicate vector control myc-tag or SMPD1-myc tag-expressing cells. ASM, acid sphingomyelinase; NPC1, Niemann–Pick type C1.