Extended Data Fig. 5. Spartin deficiency does not affect general autophagy machinery and LD biogenesis.
a, Immunoblot analyses of SUM159 wild-type and spartin KO cells. Starvation samples were incubated in serum-free and low-glucose medium (1 g/L) for 3 h prior to lysate preparation. b, Ratiometric fluorescence measurement of Keima-LC3B in SUM159 wild-type, ATG7 KO, and spartin KO in basal or amino acid depletion for 4 h. Mean ± SD, n = 14 cells (WT-Basal), 13 cells (WT-AA depletion), 10 cells (Spartin KO-Basal), 10 cells (Spartin KO-AA depletion), 11 cells (ATG7 KO-Basal), and 11 cells (ATG7 KO-AA depletion) from two independent experiments. c, Confocal imaging of fixed SUM159 wild-type and spartin KO cells stained with BODIPY493/503 and Hoechst 33342. Cells were treated with 0.5 mM OA for 30 min prior to fixation. Scale bar: 20 μm. d, Quantification of number of LDs per cell in wild-type and spartin KO SUM159 cells shown in (c) The images were taken on high-throughput confocal microscope. n = 202 cells (WT) and 240 cells (Spartin KO) from three independent experiments, median with interquartile range, n.s. not significant, two-tailed unpaired Welch’s t-test. Source numerical data and unprocessed blots are available in source data.