Fig. 1
Depletion of key autophagy factors results in mitotic centrosome fragmentation. a Abnormal mitoses in U2OS cells treated with control, ULK1 or ATG7 siRNA stained for γ-tubulin, β-tubulin, and Hoechst33342. b Time-lapse imaging of stable U2OS mRFP-α-tubulin H2B-GFP cells treated with control or ATG7 siRNA. Images were acquired every 5 min for 10 h. Time after NEBD is indicated. c Quantification of experiments represented in (b) and Supplementary Fig. 1B. Time in mitosis was measured from NEBD to anaphase or until the end of the experiment. Results are pooled from three independent experiments. siSCR, n = 89; siULK1, n = 108; siATG7, n = 119. Bars represent medians and interquartile range. *P ≤ 0.05, ****P ≤ 0.0001. Two-tailed Mann–Whitney test. d Examples of phenotype categories; bipolar, multipolar and diffuse bipolar spindles. U2OS cells are stained for γ-tubulin, β-tubulin, and Hoechst33342. e Quantification of phenotype distribution in (d). Columns represent the mean ± SD, n = 3 of ≥20 cells, ns P > 0.05, **P ≤ 0.01, ***P ≤ 0.001. Unpaired Student’s t-test, two-tailed. f Multipolar mitoses in U2OS cells depleted of ULK1 or ATG7 analyzed by β-tubulin, centrin, and Hoechst33342 staining. Numbers refer to phenotype categories in (g). White arrow, spindle pole with two centrioles; green arrow, spindle pole without centrin (PCM fragmentation); red arrow, spindle pole with abnormal centrin g. Quantification of (f). Columns represent phenotype distribution of 3 pooled experiments. siSCR, n = 7; ULK1, n = 36; ATG7 n = 37. Scale bars, 10 µm. Source data are provided as a Source Data file
