Fig. 1

AURKA directly phosphorylates ALDH1A1 and regulates its subcellular localization. a ALDH1A1 is directly phosphorylated by AURKA. AURKA-TPX2 complex was subjected to kinase assay with either [³²P]ATP (lane 1) or with 6x-His-ALDH1A1 and [³²P]ATP (lane 2) for 15 min. Lane 3 shows ALDH1A1 incubated with [³²P]ATP. b, c Subcellular localization of ALDH1A1 in (b) BxPC3 and (c) Panc1 cells treated with scrambled or ALDH1A1 shRNA for 30 h. d AURKA localization in BxPC3 and (e) Panc1 cells treated with scrambled or AURKA shRNA for 30 h. f AURKA regulates ALDH1A1 subcellular localization in BxPC3 cells. Unsynchronized cells were either treated with scrambled (top) or AURKA (bottom) shRNA for 30 h, fixed and stained with DAPI (blue) or ALDH1A1 antibody (red). More than 100 cells were analyzed from multiple random frames. Representative data are shown. g Histogram shows percentage of BxPC3 cells displaying cytosolic and perinuclear localization in response to AURKA shRNA and (i) MLN8237. Values shown are mean ± SEM of three independent experiments. * indicate statistically significant differences with respect to controls. p < 0.05 analyzed by two-way analysis of variance. h AURKA regulates ALDH1A1 subcellular localization using its kinase activity. Unsynchronized cells were treated with either DMSO or MLN8237 for 16 h, fixed and stained with DAPI or ALDH1A1 antibody. More than 100 cells were analyzed from multiple random frames. Representative data are shown. j, l AURKA ablation (j) or inhibition (l) results in somewhat perinuclear localization of ALDH1A1 in Panc1 cells. k, m Histogram shows percentage of Panc1 cells showing cytosolic and perinuclear localization in response to (k) AURKA shRNA and (m) MLN8237. Values shown are mean ± SEM of three independent experiments. * indicate statistically significant differences with respect to controls. p < 0.05 analyzed by two-way analysis of variance. Data used to generate the summary statistics shown in panels g, i, k, and m are reported in Additional file 2