(A) Quantitative analysis of Western blot analysis and fluorescence measurements (shown in Figure 1G) of the time course of in vitro PAR degradation using recombinant ARH3. Each line plot in the graph represents mean ± SEM of relative intensities (n=3). (B) Quantitative analysis of Western blot analysis and bioluminescence imaging (shown in Figure 4B) of 231-PAR-T NanoLuc cells treated with 20 µM Niraparib or 20 µM PARG inhibitor for 2 hr prior to UV radiation. Each bar in the graph represents the mean ± SEM of the relative intensities (n=3, one-way ANOVA, *p<0.05, **p<0.001, and ***p<0.0001; ns=not significant). (C) Measurements of ELISA and fluorescence intensities using 0, 0.625, 1.25, and 2.5 nM concentrations of purified PAR. Each bar in the graph in represents the mean ± SEM of the relative intensities (n=3, paired t-test, *p<0.05, **p<0.01, and ***p<0.001; ns=not significant). (D) Immunofluorescence assay using WWE-Fc to measure PAR formation in response to H2O2 using 293T cells. The cells were treated with 20 µM PJ34 (vs. untreated control, ‘Un’) for 2 hr prior to 15 min of treatment with 1 mM H2O2. The images were collected using a confocal microscope. (E) Quantification of the results in (D). Each bar in the graph represents the mean ± SEM of the relative levels of the fluorescence intensity of PAR normalized to DAPI (n=3 biological replicates with at least 150 cells in total, one-way ANOVA, ***p<0.0001). (F) Representation of the dynamic ranges of PAR-T sensors in comparison to other available PAR detection tools as indicated: (a) Western blotting with WWE-Fc versus live-cell luciferase assay using PAR-T NanoLuc was performed using UV-induced DNA damage in MDA-MB-231 Luc cells (from (B)); (b) Immunofluorescence with WWE-Fc versus live-cell imaging using PAR-T ddGFP was performed using H2O2-mediated PARP-1 activation in 293T cells (from (D)); (c) Western blotting with WWE-Fc versus fluorescence assay with PAR-T ddGFP was performed using ARH3 mediated degradation of PAR in vitro (from (A)); (d) ELISA versus fluorescence assay with PAR-T ddGFP was performed using immobilized PAR (from (C)).