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. 2019 Jan 7;218(1):251–266. doi: 10.1083/jcb.201802157

Figure 3.

Figure 3.

STRAP is a negative regulator of autophagy. (A) More JMY colocalizes with LC3 vesicles in STRAP knockout cell lines under both fed and starved conditions. (B) Quantification of colocalization of JMY and LC3 in cells either lacking STRAP or overexpressing STRAP (n = 3 independent experiments, 15 ~ 24 cells, **, P < 0.01; *, P < 0.05, Student’s t test). (C) Immunofluorescent staining of LC3 in WT and STRAP knockout cell lines. U2OS cells were either fed (top) or starved in EBSS with 100 nM Bafilomycin A to block autophagosome maturation (bottom). (D and E) Autophagosome number and size increase significantly in STRAP knockout cells (n = 3 independent experiments, 60 ∼ 68 cells, *, P < 0.05, Student’s t test). (F) Autophagic flux is increased in STRAP knockout cell lines, whereas it is decreased in JMY knockout cell lines. The autophagic flux is monitored by relative p62 protein concentrations measured by Western blot. Temperature for live cell imaging: 37°C. Scale bars: 5 µm.