Fig. 3

a Keap1-binding partners altered upon Keap1-specific-hydroxynonenylation identified by SILAC proteomics (see Supplementary Table 2, Supplementary Data 2) were knocked down using MOs targeting indicated genes in Tg(lyz:TagRFP), and Z-REX-mediated Keap1-hydroxynonenylation was performed in these knockdown backgrounds. X-axis: effect of the knockdown alone; Y-axis: the fold change in neutrophil count following Z-REX-mediated Keap1-hydroxynonenylation. These effects were not correlated. See Supplementary Fig. 9a, b. b HEK293T transfected with either empty vector or Flag-Wdr1 were treated with staurosporine for 18 h and Caspase-3 activity was measured. P values were calculated with Student’s t-test. c HEK293T transfected with either empty vector or Flag-Wdr1 were treated with staurosporine and/or Bcb for 18 h and Caspase-3 activity was measured. See Supplementary Fig. 11e, f. P values in black were calculated with Student’s t-test; P values in blue were calculated with ANOVA and Dunnett’s multiple comparisons test. d HEK293T subjected to T-REX-mediated Keap1-hydroxynonenylation and staurosporine stimulation showed Caspase-3-activity upregulation; treatment with Bcb ablated this effect. See Supplementary Fig. 11g. P values in black were calculated with Student’s t-test; P values in blue were calculated with ANOVA and Tukey’s multiple comparisons test. e HEK293T cells expressing shWdr1 or shControls were subjected to T-REX-mediated Keap1-hydroxynonenylation in the presence of staurosporine, and Caspase-3 activity was measured. See also Supplementary Fig. 12. P values in black were calculated with Student’s t-test; P values in blue were calculated with ANOVA and Tukey’s multiple comparisons test. f Antioxidant-response (AR) activity was measured in lysates from (e) to evaluate the intersection of Wdr1 with Keap1/Nrf2 signaling. Inset at right: Fold change (T-REX/light alone) in AR. P values were calculated with ANOVA and Tukey’s multiple comparisons test. g Similar experiment as in (e) to measure Caspase-3 activity in HEK293T cells expressing shCfl1 or shControls. See Supplementary Figs. 12 and 15. P values in black were calculated with Student’s t-test; P values in blue were calculated with ANOVA (see also Supplementary Fig. 1b). All data present mean ± SEM. All Student’s t-tests were two-tailed and unpaired. All sample sizes are listed in Supplementary Methods. Source data are provided as a Source data file.