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. Author manuscript; available in PMC: 2015 May 10.
Published in final edited form as: Nat Commun. 2014 Nov 10;5:5362. doi: 10.1038/ncomms6362

Figure 7. PARP13 depletion results in resistance to TRAIL-mediated apoptosis.

Figure 7

a, Percent survival of untreated and TRAIL treated wild type cells and wild type cells expressing TRAILR4-Flag assayed via Annexin-V/PI FACS (average of n=3 independent experiments, bars represent SEM, p<0.05(*), two-sided t-test). b, Immunoblot of TRAILR1-2 and TRAILR4 proteins in wild type and PARP13−/−A cells. GAPDH is used as loading control. c, Survival assay measuring proliferation of SW480, HCT116 and HeLa cells with or without PARP13 knockdown after treatment with increasing concentrations of TRAIL for 24 h. Results are shown relative to untreated cells (means of n=3 independent experiments, error bars show SEM). Results for double knockdown of TRAILR4 and PARP13 are shown for HeLa cells. d, TRAILR4 mRNA levels after PARP13 and PARP13+TRAILR4 knockdown relative to control knockdown (averages of n=3 independent experiments, error bars show SD). e, Annexin-V/PI apoptosis assays comparing the percent survival of wild type and three independent PARP13−/− cell lines (A, B, C) upon 1 µg/ml TRAIL treatment for 24 h (n=3 independent experiments, bars show SEM, asterisks show significance relative to wild type, p<0.001 (***), two-sided t-test) f, Normalized survival of wild type and three PARP13−/− cell lines treated with 1µg/ml TRAIL relative to untreated (averages of n=3 independent experiments, bars show SEM, asterisks show significance relative to wild type, p<0.05 (*) or p<0.01(**), two-sided t-test). g, Colony formation assay measured by crystal violet staining of wild type or PARP13−/−A cells treated with or without the indicated amounts of TRAIL for 7 days. h, Annexin-V/PI apoptosis assays comparing the percent survival of wild type, PARP13−/−A or PARP13−/−A cells expressing PARP13.1, PARP13.1VYFHR or PARP13.1ΔZnF upon treatment with or without 1 µg/ml TRAIL for 24 h. Data is shown as % survival (means of n=3 independent experiments, bars show SEM, asterisks show significance relative to wild type, p<0.05 (*), p<0.01(**), p>0.05 (n.s.), two-sided t-test). i, Normalized survival of wild type, PARP13−/−A cells and PARP13−/−A cells expressing PARP13.1, PARP13.1VYFHR or PARP13.1ΔZnF treated with 1µg/ml TRAIL relative to untreated (n=3 independent experiments, bars show SEM, asterisks represent significance relative to wild type, p<0.05 (*), p<0.01(**), p<0.001(***), two-sided t-test).