**Appendix C**

*Int. J. Mol. Sci.* **2020**, *21*, x FOR PEER REVIEW 17 of 23

**Figure A2.** Concentrations of 150 nM OLA and BLEO. A single experiment was carried out to explore

evaluated using MTT (72 h). Mean ± SD. *n* = 6. Since the predicted additive effect according to Webb's

equation [25] was equal or lower than the combined effect, synergism was discarded.

**Figure A3.** PARPis and BLEO in VERO. PARPis and BLEO in VERO. (**A**) Scheme representing PAR, its synthesis by PARPs, its degradation mainly by poly-ADP-glycohydrolase (PARG) activity, and the inhibitors abbreviations associated with the correspondent enzyme. 3AB, EB, and OLA are PARPis, while DEA inhibits PARG. (**B**) Test of the inhibitors' effects on PAR. The inhibitors used were 3 aminobenzamide (3AB, Sigma A-0788), 5′-deoxy-5′-[4-[2-[(2,3-dihydro-1oxo-1H-isoindol-4 yl)amino]-2-oxoethyl]-1-piperazinyl]-5′-oxoadenosine dihydrochloride (EB, Alexis Biochemicals), 6,9-diamino-2-ethoxyacridine-DL-lactate monohydrate (DEA, Trevigen 4680-096-03), and OLA. PAR was measured on confocal planes of monolayers using the Analyze/Measure ImageJ plugin, and the results are expressed as arbitrary fluorescence units/area. (**C**–**F**) Cell viability assays. Results are expressed as % MTT absorbance with respect to control. The violet arrows at the top of each graph represent the time in days (5 days) from seeding to MTT evaluation. The bleomycin treatment (40 to 80 µg/mL, 45 min) is represented in green and the inhibitors' treatment durations are in red. (**C**) Pretreatment with 3AB or EB, (**D**) co-treatment with 3AB or EB, (**E**) pre- and co-treatment with the PARG **Figure A3.** PARPis and BLEO in VERO. PARPis and BLEO in VERO. (**A**) Scheme representing PAR, its synthesis by PARPs, its degradation mainly by poly-ADP-glycohydrolase (PARG) activity, and the inhibitors abbreviations associated with the correspondent enzyme. 3AB, EB, and OLA are PARPis, while DEA inhibits PARG. (**B**) Test of the inhibitors' effects on PAR. The inhibitors used were 3-aminobenzamide (3AB, Sigma A-0788), 50 -deoxy-50 -[4-[2-[(2,3-dihydro-1oxo-1H-isoindol-4-yl)amino]- 2-oxoethyl]-1-piperazinyl]-50 -oxoadenosine dihydrochloride (EB, Alexis Biochemicals), 6,9-diamino-2-ethoxyacridine-DL-lactate monohydrate (DEA, Trevigen 4680-096-03), and OLA. PAR was measured on confocal planes of monolayers using the Analyze/Measure ImageJ plugin, and the results are expressed as arbitrary fluorescence units/area. (**C**–**F**) Cell viability assays. Results are expressed as % MTT absorbance with respect to control. The violet arrows at the top of each graph represent the time in days (5 days) from seeding to MTT evaluation. The bleomycin treatment (40 to 80 µg/mL, 45 min) is represented in green and the inhibitors' treatment durations are in red. (**C**) Pre-treatment with 3AB or EB, (**D**) co-treatment with 3AB or EB, (**E**) pre- and co-treatment with the PARG inhibitor DEA, and (**F**) co- and post-treatment with DEA. The genotoxic treatment was always different from the control (*p* < 0.001). Asterisks refer to a comparison of a treatment vs. the control or with respect to the genotoxic treatment. \*: *p* < 0.05, \*\*: *p* < 0.01. *Int. J. Mol. Sci.* **2020**, *21*, x FOR PEER REVIEW 18 of 23 inhibitor DEA, and (**F**) co- and post-treatment with DEA. The genotoxic treatment was always
