Journal: The EMBO Journal

Article Title: Parp7 generates an ADP-ribosyl degron that controls negative feedback of androgen signaling

doi: 10.1038/s44318-025-00510-4

Figure Lengend Snippet: ( A – C ) Immunoblot detection of AR and AR-ADPr (by FL-AF1521) in VCaP cells subjected to a time course of R1881 ( A ), R1881 and RBN2397 ( B ), or R1881 and Bortezomib (Bortez) (C) treatment. Cells were collected every 2 h for an 18-h period. ( D ) Line plot visualizing the AR protein density measurements for immunoblots from ( A – C ). The y axis represents the log 2 of the AR/TUB density fold change from 0-h time point, and the x axis represents the time of treatment in hours. The curves were fitted using the loess method. Data points represent single measurements. ( E ) Schematic diagram of the androgen-induced AR degradation mechanism. ( F ) Immunoblot detection of AR and AR-ADPr (Fl-AF1521) in VCaP cells treated with DHT, DHT + RBN2397, and DHT+Bortezomib for times indicated on the panel. ( G ) Immunoblot detection of the AR and AR-V7 protein in VCaP cell extracts and AF1521 bound fractions. Cell extracts from VCaP cells treated with different combinations of R1881, RBN2397, and Bortezomib for 10 h and were combined with AF1521 beads for the enrichment of ADP-ribosylated proteins. ( H ) Immunoblot detection of Flag-AR, AR-ADPr, and FKBP51 in PC3-AR shCTRL (left) and shPARP7 (right) cells treated with R1881 and cotreated with R1881 and RBN2397 for times indicated on the panel. The shGFP was used as shCTRL. ( I ) Immunoblot detection of Flag-AR and AR-ADPr in PC3-AR cells treated for 18 h with R1881 and different PARP inhibitors (RBN2397—PARP7i; RBN012759—PARP14i; Veliparib, Olaparib, and Talazoparib—PARP1/2i). .

Article Snippet: Synthetic androgen R1881 (Perkin-Elmer, NLP005005MG; used at 2 nM final concentration in most experiments), natural androgen agonist dihydrotestosterone (DHT, Sigma-Aldrich, D-073), proteasome inhibitor Bortezomib (MedChemExpress, HY-10227; used at 1 μM final concentration), PARP7 inhibitor RBN2397 (DC Chemicals, DC31069), PARP14 inhibitor RBN012759 (MedChemExpress, HY-136979), and PARP1/2 inhibitors Veliparib (Selleck Chemical LLC, S100410MG), Olaparib (MedChemExpress, HY-10162), and Talazoparib (MedChemExpress, HY-16106) were used at concentrations and treatment times specified in the figures or figure legends.

Techniques: Western Blot

Journal: Molecular Cell

Article Title: Mitochondrial NAD + Controls Nuclear ARTD1-Induced ADP-Ribosylation

doi: 10.1016/j.molcel.2020.12.034

Figure Lengend Snippet:

Article Snippet: Olaparib , Selleckchem , Cat# S1060.

Techniques: Recombinant, Modification, Reverse Transcription

Journal: Journal of Clinical Medicine

Article Title: Low-Dose and Long-Term Olaparib Treatment Sensitizes MDA-MB-231 and SUM1315 Triple-Negative Breast Cancers Spheroids to Fractioned Radiotherapy

doi: 10.3390/jcm9010064

Figure Lengend Snippet: DNA Double Strand Breaks (DSBs) induction in MDA-MB-231 ( A ) and SUM1315 ( B ) cell lines cultured in monolayer after a treatment with 0.1% DMSO (control cells), 5 and 50 µM Olaparib for 4 h alone (no irradiation “no RX” ) or combined with 2 Gy of irradiation (4 h and 2 Gy). The number of foci gammaH2AX per cell in >10 fields per condition was determined with ImageJ software. Results are represented as mean number of foci gammaH2AX per nuclei with their standard deviation. A student t -test was performed for statistical comparison of all treatment conditions, where ns = non-significant and ***** p < 0.00001. Magnification = 40×, scale bar = 50 µm.

Article Snippet: Olaparib (Carbosynth ® ) was solubilized in Dimethylsulfoxide (DMSO) at a concentration of 100 mM.

Techniques: Cell Culture, Control, Irradiation, Software, Standard Deviation, Comparison

Journal: Journal of Clinical Medicine

Article Title: Low-Dose and Long-Term Olaparib Treatment Sensitizes MDA-MB-231 and SUM1315 Triple-Negative Breast Cancers Spheroids to Fractioned Radiotherapy

doi: 10.3390/jcm9010064

Figure Lengend Snippet: Clonogenic cell survival of MDA-MB-231 ( A ) and SUM1315 ( B ) cell lines cultured in monolayer, after a treatment with 0.5, 5 and 50 µM Olaparib alone for 24 h, 72 h, or 120 h ( a ), or combined with fractioned irradiation of 2, 6 or 10 Gy (“Rx dose”) ( b ). For each treatment condition, the percentage of the clonogenic survival was determined by the ratio of the survival fraction of treated cells on the survival fraction of 0.1% DMSO control cells. Results are represented on a Heatmap. Standard errors are <10% for every treatment condition ( n > 3). ( c ) Photographs of clonogenic populations of MDA-MB-231 and SUM1315, Magnification = 40×, scale bar = 50 µm.

Article Snippet: Olaparib (Carbosynth ® ) was solubilized in Dimethylsulfoxide (DMSO) at a concentration of 100 mM.

Techniques: Cell Culture, Irradiation, Control

Journal: Journal of Clinical Medicine

Article Title: Low-Dose and Long-Term Olaparib Treatment Sensitizes MDA-MB-231 and SUM1315 Triple-Negative Breast Cancers Spheroids to Fractioned Radiotherapy

doi: 10.3390/jcm9010064

Figure Lengend Snippet: MDA-MB-231 spheroids live/dead profile ( A – C,G – I ) and metabolic activity ( D – F ) after a treatment with Olaparib (“Ola”, 5 and 5 µM) for 6, 8 and 10 days with or without fractioned irradiation (“Rx”). ( A – C,G – I ) Spheroid viability/mortality profile and spheroid normalized size after a treatment with Olaparib and/or fractioned irradiation for 6 days ( A , G ), 8 days ( B , H ) and 10 days ( C , I ): Images were taken with Cytation3MV where green markings correspond to calcein-AM penetration (viable cells) and red markings correspond to ethidium homodimer-1 cell penetration (dead cells). The percentage of spheroid size was normalized by the size of spheroid controls. Scale bar = 200 µm. ( D – F ) MDA-MB-231 cell metabolic activity in spheroids with resazurin test after 6 ( D ), 8 ( E ) and 10 days of treatment ( F ). Corrected fluorescence intensity of resorufin (ex/em λ576/λ584 nm) was measured after 15 h incubation with 60 µM resazurin. The fluorescence intensity of treated spheroids ( n > 15) was normalized by the fluorescence intensity of each associated 0.1% DMSO control spheroids. Results are expressed as mean metabolic activity (%) of each treatment conditions with their standard errors. A student t -test was performed for the statistical comparison of all treatment conditions and the p -value is represented in the graphs as ns = non-significant, ** p < 0.01 and ***** p < 0.00001.

Article Snippet: Olaparib (Carbosynth ® ) was solubilized in Dimethylsulfoxide (DMSO) at a concentration of 100 mM.

Techniques: Activity Assay, Irradiation, Fluorescence, Incubation, Control, Comparison

Journal: Journal of Clinical Medicine

Article Title: Low-Dose and Long-Term Olaparib Treatment Sensitizes MDA-MB-231 and SUM1315 Triple-Negative Breast Cancers Spheroids to Fractioned Radiotherapy

doi: 10.3390/jcm9010064

Figure Lengend Snippet: SUM1315 spheroids live/dead profile ( A – C,G – I ) and metabolic activity ( D – F ) after a treatment with Olaparib (“Ola”, 5 and 5 µM) for 6, 8 and 10 days with or without fractioned irradiation (“Rx”). ( A – C,G – I ) Spheroid viability/mortality profile and spheroid normalized size after a treatment with Olaparib and/or fractioned irradiation for 6 days ( A , G ), 8 days ( B , H ) and 10 days ( C , I ): Images were taken with Cytation3MV where green markings correspond to calcein-AM penetration (viable cells) and red markings correspond to ethidium homodimer-1 cell penetration (dead cells). The percentage of spheroid size was normalized by the size of spheroid controls. Scale bar = 200 µm. ( D – F ) SUM1315 cell metabolic activity in spheroids with resazurin test after 6 ( D ), 8 ( E ) and 10 days of treatment ( F ). Corrected fluorescence intensity of resorufin (ex/em λ576/λ584 nm) was measured after 15 h incubation with 60 µM resazurin. The fluorescence intensity of treated spheroids ( n > 15) was normalized by the fluorescence intensity of each associated 0.1% DMSO control spheroids. Results are expressed as mean metabolic activity (%) of each treatment conditions with their standard errors. A student t-test was performed for the statistical comparison of all treatment conditions and the p value is represented in the graphs as ns = non-significant, * p < 0.05, ** p < 0.01, *** p < 0.001 and ***** p < 0.00001.

Article Snippet: Olaparib (Carbosynth ® ) was solubilized in Dimethylsulfoxide (DMSO) at a concentration of 100 mM.

Techniques: Activity Assay, Irradiation, Fluorescence, Incubation, Control, Comparison