Fig. 4: Arachidonic acid-derived PROTAC degraded FTH1 in the presence of MOF-Fe.

a Schemes of PROTAC library. b Chemical structure of C20U4V. c Scattered plots of the IC₅₀ values of the PROTAC library against Panc-1 and MIAPaCa-2 cells after 72 h treatment, n = 6 independent experiments. d Scatter plots of the change of FTH1 expression in Panc-1 cells after treatment with MOF-Fe (3 μg Fe/mL) and the PROTAC library for 12 h, n = 3 independent experiments. e Immunoblots of FTH1 degradation by C20U4V in the presence of MOF-Fe, n = 3 independent experiments. Quantification is included in Supplementary Fig. 7e. f Volcano plots of protein expression in Panc-1 cells treated for 12 h with MOF-Fe + C20U4V vs. MOF-Fe. Differentially expressed proteins in the ferroptosis pathway are labeled. MOF-Fe (3 μg Fe/mL), C20U4V (20 μM), n = 3 independent experiments. g Chemical structure of BPAA and immunoblots of FTH1 pulled down by BPAA. The samples derived from the same experiments but different gels were processed in parallel: FTH1 pulldown was detected on one gel, and the input controls of FTH1 and β-actin were run on another gel, n = 3 independent experiments. Quantification is included in Supplementary Fig. 8a. h Immunoblots of HA-tagged ubiquitin (HA-Ub) immunoprecipitated by anti-Flag antibody in the lysate of HEK-293T. The HEK-293T cells were transfected with HA-Ub and Flag-FTH1 plasmids, and treated with C20U4V (10 μM) in the presence of MG132 (10 μM) for 6 h. The samples derived from the same experiment but different gels were processed in parallel: IP detection of HA-Ub and Flag-FTH1 were run on one gel, and the input controls of HA-Ub, Flag-FTH1, and β-actin were run on another gel. The experiments were independently repeated twice with similar results. A biologically independent replicate is shown in Supplementary Fig. 9a. i FTH1 expression after incubation with MOF-Fe and AA for 48 h, n = 3 independent experiments. Quantification is included in Supplementary Fig. 9b. j FTH1 expression after incubation with MOF-Fe and (S,S,S)-C20U4V for 12 h. Quantification is included in Supplementary Fig. 9c, n = 3 independent experiments. k VHL expression in VHL-knockdown Panc-1 cells. The experiments were independently repeated twice with similar results. l FTH1 expression in NC or shVHL-knockdown cells after incubation with MOF-Fe and C20U4V for 48 h. Quantification is included in Supplementary Fig. 9d, n = 3 independent experiments. Data were presented as mean ± SEM. Statistical significance was determined using multiple one-tailed ratio-paired t-test without correction for multiple comparison (d) or EdgeR analysis using negative binomial generalized linear model (GLM) adjusted with Benjamini–Hochberg algorithm (f), and p values are indicated. Source data are provided as a Source Data file.