Extended Data Fig. 4: Spartin-mediated lipophagy is ubiquitination-independent.
From: The Troyer syndrome protein spartin mediates selective autophagy of lipid droplets
a. Immunoblot analysis of SUM159 cells wild-type and cells lacking ATG7 with the indicated antibodies. b, Immunoblot analysis of SUM159 wild-type cells with the indicated antibodies. Cells were treated with DMSO (vehicle) or 2 μM TAK-243 for 2 h prior to cell lysis. c, Confocal imaging of live SUM159 cells transiently expressing LAMP1–mNG and mCherry–LiveDrop. Cells were treated with 0.5 mM OA for 24 h and chased for 3 h after OA withdrawal, with DMSO (vehicle) or 2 μM TAK-243 for 2 h prior to imaging. Scale bars: full-size, 20 μm; insets, 2 μm. d, Overlap between LiveDrop and LAMP1 in cell periphery shown in (c) was quantified by Pearson’s coefficient analysis. Mean ± SD, n = 10 cells from two independent experiments, n.s. not significant, two-tailed unpaired t-test. e, Immunoblot analyses of SUM159 wild-type cells with indicated antibodies. Cells were treated with 20 nM negative control siRNA or selective autophagy receptors (SARs; OPTN, NBR1, SQSTM1/p62) siRNAs for 72 h prior to lysate preparation. f, Ratiometric fluorescence measurement of Keima–LiveDrop in SUM159 wild-type cells treated with control siRNA (iNC) or SARs siRNAs for 72 h, n.s. not significant, two-tailed unpaired t-test. Mean ± SD, n = 17 cells from three independent experiments. Source numerical data and unprocessed blots are available in source data.
