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. 2021 Apr 30;220(6):e202103105. doi: 10.1083/jcb.202103105

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© 2021 Li et al.

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Figure 4. — TMEM41B and VMP1 have scramblase activity in vitro. (A) Schematic of the fluorescence-based scrambling assay. Only the NBD-lipids in the outer leaflet of liposome can be bleached into nonfluorescent ABD-lipid by dithionite. (B) Scrambling of NBD-lipids by TMEM41B (top), VMP1 (middle), and TMEM41B-VMP1 complex (bottom) with different concentrations of proteins as indicated. The time points of adding dithionite and Tween 20 are denoted. Data represent the average of three independent measurements using the same batch of NBD-lipid–containing liposomes. (C–E) Substrate preference for TMEM41B (left) and VMP1 (right) against NBD-PC (C), NBD-PE (D), and NBD-PS (E). The concentration of tested protein is indicated. The time points of adding dithionite and Tween 20 are denoted. Data represent the average of three independent measurements using the same batch of NBD-lipid–containing liposomes.