Fig. 4
ADSL is hydroxylated by EglN2 on Proline 24. a In vitro hydroxylation of GST-ADSL in the presence or absence of recombinant EglN2. Bar graph represents the normalized ratio of the intensity of the oxidized P24-containing peptide to that of ADSL full protein. Error bars represent SEM, n = 4, *P < 0.05 was calculated using two-tailed Student’s t-test. b Fragmentation spectrum of proline hydroxylated peptide YASPEMCFVFSDR detected following experimental procedure described in (a). c, d IB of WCE and IP of lysates from 293T cells transfected with the indicated plasmids, and treated as indicated overnight. e IB of lysates from MDA-MB-231 cells overexpressing the indicated dox-inducible ADSL, transduced and treated as indicated. f Representative image of 2-D proliferation of MDA-MB-231 cells overexpressing the indicated dox-inducible ADSL, transduced and treated as indicated. g Enzymatic activity of WT, P24A or R426H ADSL in the presence of its substrate adenylosuccinate (SAMP) or succinylaminoimidazole carboxamide ribotide (SAICAR). Bar graphs represent the normalized percentage of ADSL activity compared with WT ADSL. Error bars represent SEM, n = 4, *P < 0.05, **P < 0.01, ***P < 0.001 were calculated using one-way ANOVA followed by Tukey’s multiple comparison test. Source data are provided as a Source Data file
