In brief, 30 g of protein per sample was resolved on SDS/PAGE gels and transferred to Immobilon-P membranes (EMD Millipore)

In brief, 30 g of protein per sample was resolved on SDS/PAGE gels and transferred to Immobilon-P membranes (EMD Millipore). that a disease with an DRI-C21045 identical genetic DRI-C21045 makeup as its human counterpart could be established in the animals. The expression of red fluorescent protein (RFP) is linked to that of to track the progeny cells derived from the transplanted HPCs (Fig. 2and Fig. S1 0.05). These data demonstrate that our animal model accurately recapitulates the physiopathology and genetic makeup of human DLBCL, particularly the high PARP1 levels in the disease. Open in a separate window Fig. 2. A mouse DLBCL model recapitulates the pathophysiology of human DLBCL. (= 5) are larger than those of B6 mice (= 5), and that most cells express RFP. (= 5) compared with B6 mice (= 5). Error bars represent SEM. * 0.05, **** 0.0001, nonparametric Students test. Open in a separate window Fig. S1. Physiological characteristics of the DLBCL mice. ( 0.01; **** 0.0001, two-tailed TSPAN8 Students test with assumption of unequal SD. Evaluation of PARP1 as a DLBCL Diagnostic Marker. To test the feasibility of PARP1 imaging in DLBCL, we used a fluorescent PARP1-targeted imaging probe, PARPi-FL (25C27). PARPi-FL is based on the PARP-specific inhibitor olaparib, a Food and Drug Administration-approved drug for treating ovarian, lung, and breast cancer with mutations (28) (Fig. 3= DRI-C21045 5) and B6 mice (= 5). The concentration of PARPi-FL is presented as the percentage DRI-C21045 of injected dose per gram of tissue (%ID/g). ( 0.05; ** 0.01; *** 0.001; **** 0.0001, nonparametric Students test. Open in a separate window Fig. S2. Confocal microscopic images of DLBCL and B6 mice. ( 0.001) (Fig. 4= 7 for [18F]PARPi; = 4 for PARPi-FL), demonstrating similar specificity of the two PARP1-targeted probes. (= 7), B6 mice (= 4), and DLBCL mice preexposed to olaparib (= 4). (= 7). Pearson correlation was used to calculate statistics and correlation coefficients. (= 9) and B6 mice (= 4) as measured by ex vivo -counting. Error bars represent SEM. * 0.05; ** 0.01; *** 0.001; **** 0.0001, nonparametric Students test. Open in a separate window Fig. S3. PET/CT imaging of B6 mice and key parameters of single-cell radioactivity measurements in DLBCL mouse blood. (= 4) and preblocked with olaparib (= 3). ( 0.01, two-tailed Students test with assumption of unequal SD. PARP1-Targeted PET Imaging for DLBCL Diagnosis. PET/CT imaging is the current standard for diagnosing and monitoring DLBCL (34). To test whether [18F]PARPi is a feasible PET imaging probe for DLBCL, we first measured its specificity in our DLBCL mouse model. We performed [18F]PARPi PET/CT imaging (Fig. 4= 7), B6 mice (= 4), and DLBCL mice preinjected with olaparib (500 g, 1.15 mol; = 4). Compared with B6 mice, DLBCL mice exhibited a 5.6-fold increase in PET signal in their lymph nodes (= 0.002). Preinjection of olaparib decreased the accumulation by 87% (= 0.0012), demonstrating DRI-C21045 that [18F]PARPi accumulation was specific to PARP1 (Fig. 4 and and = 0.0028) (Fig. 4and and = 12), B6 mice injected with [18F]PARPi (= 4), and DLBCL mice injected with [18F]PARPi and blocked with olaparib (= 7). (= 4) or preblocked with olaparib (= 3). (= 5) and B6 mice (= 5). Consecutive sections were stained with H&E, PARP1, or RFP. Error bars represent SD. * 0.05; ** 0.01; *** 0.001; **** 0.0001, nonparametric Students test. PARP1-Targeted PET Imaging to Differentiate Malignant from Inflamed Lymph Nodes. Because of the high uptake of glucose exhibited by inflammatory immune cells in.