Supplementary Materials and Methods, Supplementary Tables 1 through 3, and Supplementary Figure 1 from The PARP Inhibitor AZD2461 Provides Insights into the Role of PARP3 Inhibition for Both Synthetic Lethality and Tolerability with Chemotherapy in Preclinical Models

Supplementary materials and methods. Suppl Table 1. MMS combination PF50 cell line assays. Concentration-dependent potentiation of MMS (PF50 ratios). Suppl Table 2. Permeability of AZD2461 and olaparib were assessed. Suppl Table 3. Activity of olaparib and AZD2461 alone or in combination with the P-gp inhibitor verapamil in the colorectal cancer cell line HCT-15, which expresses high levels of P-gp. Suppl Fig. 1. Immunofluorescence for poly(ADP-ribose) in human A549 cells. Suppl Fig. 2. Analysis of both in vivo pharmacokinetics (PK) and pharmacodynamics (PD) in tumors taken from SW620 (human colon carcinoma) xenograft-bearing mice. Suppl Fig. 3. Colony formation (clonogenic) assays in the intrinsically high P-gp-expressing MRE11-deficient HCT-15 human colorectal cell line. Suppl. Fig. 4. Combination efficacy studies. Suppl Fig. 5. Relative body weight changes in mice following treatment with 10 mg/kg AZD2461 and olaparib in combination with temozolomide. Suppl. Fig. 6. A serial dilution of RNA concentration was used to establish a standard curve for determining the reaction efficiency of PARP1, PARP2, PARP3, and housekeeper gene (PPIA: cyclophilin A) TaqMan RT-PCR assays in both rat and mouse species; Determination of optimal reference gene for rat-mouse data normalization. Suppl Fig. 7.A. A serial dilution of reference RNA (Qiagen) concentration was used to establish a standard curve for determining the reaction efficiency of PARP3 and housekeeper genes (PPIA: cyclophilin and YWHAZ: tyrosine 3-monooxygenase/tryptohpan 5-monooxygenase activation protein, zeta polypeptide) SybrGreen RT-PCR assays in both rat and human species; Determination of optimal reference gene for rat-human data normalization. Suppl Fig. 8. Relative body weight changes in rat following treatment with 10 mg/kg AZD2461 and olaparib in combination with temozolomide; Relative body weight changes in rat following treatment with 20 mg/kg AZD2461 and olaparib in combination with temozolomide.