Fig. 1: PEF upregulated ferroptosis suppressor genes in pancreatic cancer cells.

a Flow cytometry histograms and quantification of C11-BODIPY oxidation, n = 7 independent experiments. b MDA concentration, n = 3 independent experiments. c Experiment scheme for panel (d). d Relative Panc-1 viability after PEF and incubation with Lip-1, Ferr-1, or tocopherol, n = 6 independent experiments. e GO analysis of biological process (GO_BP) for genes upregulated in the PEF +  cell debris group relative to the vehicle control group. f Volcano plots of transcriptome. Differentially expressed genes (DEGs) in the ferroptosis pathway are labeled with red (upregulated) or blue (downregulated). g GSEA on the WikiPathway gene set. h Volcano plots of transcriptome. DEGs in the NRF2 pathway are labeled with red (upregulated) or blue (downregulated). Cells in panels e–h were treated with PEF and incubated with cell debris for 24 h, n = 3 independent experiments. i Immunoblots of Panc-1 cells. The samples derived from the same experiment but on different gels for NRF2, another for HMOX1, FTH1, and β-actin, were processed in parallel, n = 3 (NRF2), 4 (HMOX1), and 5 (FTH1) independent experiments. Quantification is included in Supplementary Fig. 1f. j Hematoxylin-eosin (H&E) staining of KPC-A719 tumors at 24 h after PEF ablation. The border between the viable tumors and the ablation zone is marked with a black dashed line. Residual cancer cells are indicated with black arrowheads. The experiments were independently repeated three times with similar results. Scale bar = 100 μm. k 4-HNE staining of KPC-A719 tumors and quantification, n = 8 randomly selected field-of-views (FOVs) from three independent mice per group. Scale bar = 100 μm. l Immunoblots of KPC-A719 tumors. The samples derived from the same experiment but on different gels for NRF2, another for HMOX1, FTH1, and β-actin, were processed in parallel, n = 9 (NRF2, HMOX1) or 12 (FTH1) biological replicates from three independent mice per time point. Quantification is included in Supplementary Fig. 1g. PEF intensity = 600 V/cm in cell culture or as specified for in vivo studies. Data were presented as mean ± standard error of mean (SEM). Statistical significance was determined using one-tailed ratio-paired t-test (a), one-tailed unpaired t-test (b, k), one-way ANOVA with Dunnett’s multiple comparison (d), GO analysis adjusted with Benjamini–Hochberg algorithm (e), EdgeR analysis using negative binomial generalized linear model (GLM) (f, h) or GSEA analysis (g) adjusted with Benjamini–Hochberg algorithm, and p values are indicated. Source data are provided as a Source Data file. The elements in Fig. 1c were generated using PowerPoint.