Supplementary Figures from Targeting NAD<sup>+</sup>/PARP DNA Repair Pathway as a Novel Therapeutic Approach to <i>SDHB</i>-Mutated Cluster I Pheochromocytoma and Paraganglioma

Supplementary Figures S1 A. Quantitative PCR showed CA9, PNMT, and VEGFA expression in MTT cells. Higher expressions of hypoxia related genes were detected in SDHBKD cells. B. NAD+ quantification confirmed depletion of NAD+ by Nampt inhibitors FK866 and CHS828. C. Blue native gel analysis on mitochondrial complex formation in MTT cells (left panel). Mitochondrial complex I formation is increased in SDHBKD cells. In gel enzyme activity assay (IGA) showed elevated mitochondrial complex I activity in SDHBKD cells (right panel). D. Re-expression of SDHB in MTT cells (upper panel). The re-expression of SDHB reduced pADPR formation in SDHBKD cells, with a corresponding increased of γH2A.X expression (lower panel). E. Quantification of NAD+/NADH ratio in MTT cells. Re-expression of SDHB partially reversed the elevated NAD+/NADH ratio in SDHBKD cells. F. Quantification of flow cytometry data. A significant G2/M arrest is seen in combination treated group. G. Western blot for MGMT detection in MTT cells. No MGMT expression was seen in either SDHBWT or SDHBKD cells.ï€ ï�¢-actin was used as loading control. H. Cell viability test showing enhanced cytotoxic effect of combination treatment. **p<0.01. I. In vivo bioluminescence showing significantly reduced metastatic lesion with TMZ and combination treatments. *p<0.05, **p<0.01.