Unbiased profiling reveals concomitant ER stress and oxidative dysregulation
Table S1: PANC-1 cells were treated with 2a or DMSO for 12 hours under hypoxia after which metabolites were extracted and identified by LC-qTOF-MS. Statistically significant changes (p < 0.05, VIP > 1) in metabolite levels are plotted relative to DMSO-treated cells. N = 10. Figure 3c: GSH-depletion in PANC-1 cells by APD-CLDs were confirmed by GSH-GloTM assay (Promega) after 12 hours under hypoxia or normoxia. Sidak’s multiple comparisons test. N = 2. Likewise, the ROS levels was quantified by fluorescence of the ROS-activated fluorophore H2DCFDA (Molecular Probes) in normoxic versus hypoxic PANC-1 cells treated with 1. Data is normalized to the average of the two lowest dosings of rakicidin A. N = 3. Unpaired t-test. Figure 3d + GSEA: PANC-1 cells were treated as in described for table S1 and after 12 hours total RNA was isolated. Transcripts were sequenced, quantified and the combined gene expression of 1 and 2a-treated cells was plotted relative to DMSO-treated cells. N = 3. See full RNAseq dataset via link below. Figure 3f: RT-qPCR on CHAC1 transcription after treatment with erastin, 1 and 2b for 12 hours under hypoxia or normoxia. All data is normalized to normoxic DMSO treatments. Sidak’s multiple comparisons test (hypoxia vs. normoxia). N = 3.