Supplementary Figures S1-S5 and Tables S1-S10, S15-S23 from Isoflavone ME-344 Disrupts Redox Homeostasis and Mitochondrial Function by Targeting Heme Oxygenase 1

Abstract

<p>Supplementary Figure S1. Effects of HO-1 knockdown in H460 cell lines and HO-1 over-expression in H596 cell lines. Supplementary Figure S2. Affinity Enrichment Mass Spectrometry. Supplementary Figure S3. ME-344 induced apoptosis in sensitive cell lines H460 and SHP-77 through caspase 3 activation. Supplementary Figure S4. NAC attenuated ME-344-induced increases in ROS levels in H460 and H596 cell lines. Supplementary Figure S5. ME-344-induced changes in protein expression of VDAC1 and VDAC2 in drug sensitive (H460 and SHP-77), resistant (H596 and SW900) or normal cell lines (MRC-5). Supplementary Table S1. The immunohistochemistry score system for human lung tissue microarrays (TMAs). Supplementary Table S2. Median fluorescent intensity fold changes of ROS in sensitive and resistant human lung cancer cell lines and human normal lung cell line after treatment of ME-344 (relative to control). Supplementary Table S3. Relative protein basal level of Nrf2, HO-1, GSTP, ALDH in sensitive, resistant and normal cell lines (Protein/β-actin, relative to H460). Supplementary Table S4. Relative mRNA basal level of Nrf2, HO-1, GSTP, ALDH in sensitive, resistant and normal cell lines (mRNA/18S rRNA, relative to H460). Supplementary Table S5. Relative protein fold change of Nrf2, HO-1, GSTP, ALDH in sensitive, resistant and normal cell lines after treatment of ME-344 (Protein/β-actin, relative to control). Supplementary Table S6. Relative mRNA fold change of Nrf2, HO-1, GSTP, ALDH in sensitive, resistant and normal cell lines after treatment of ME-344 (mRNA/18S rRNA, relative to control). Supplementary Table S7. Relative mRNA basal level of UPR markers in sensitive, resistant and normal cell lines (mRNA/18S rRNA, relative to H460). Supplementary Table S8. Relative mRNA fold change of UPR markers in sensitive, resistant and normal cell lines after treatment of ME-344 (mRNA/18S rRNA, relative to control). Supplementary Table S9. Relative protein basal level of UPR markers in sensitive, resistant and normal cell lines (protein/β-actin, relative to H460). Supplementary Table S10. Relative protein fold change of UPR markers in sensitive, resistant and normal cell lines after treatment of ME-344 (protein/β-actin, relative to control). Supplementary Table S15. Immunohistochemistry scores of HO-1 in lung normal and tumor specimens of different histological types. Supplementary Table S16. Immunohistochemistry scores of Nrf2 in lung normal and tumor specimens of different histological types. Supplementary Table S17. Immunohistochemistry scores of HO-1 and Nrf2 in lung normal and tumor specimens of different staging. Supplementary Table S18. GSH (nmol/mg protein) generation of H460 and H596 cell lines with pretreatment of NAC. Supplementary Table S19. IC50 (µM) of H460 and H596 cell lines with and without pretreatment of NAC. Supplementary Table S20. Relative protein basal level of VDAC1 and VDAC2 in sensitive, resistant and normal cell lines (Protein/β-actin, relative to H460). Supplementary Table S21. Relative protein fold change of VDAC1 and VDAC2 in sensitive, resistant and normal cell lines after treatment of ME-344 (Protein/β-actin, relative to control). Supplementary Table S22. Basal and fold change after ME-344 treatment of ROS levels and protein expression of Nrf2 and HO-1. Supplementary Table S23. Basal and fold change after ME-344 treatment of ROS levels in HO-1 knockdown and overexpressing H460 and H596 cell lines.</p>