Journal: Journal of cell science

Article Title: Cancer metabolism in radiation sensitization: complementary roles of O-GlcNAc transferase (OGT) and PARP1

doi: 10.1242/jcs.264322

Figure Lengend Snippet: A . MCF7 cells were treated for 1 h with DMSO or 50 μM PUGNAc ± 10 μM veliparib, irradiated with 6 Gy, fixed after 2 h and stained for 53BP1 and BRCA1. Representative images with 53BP1 foci (green), BRCA1 foci (red), and DAPI (blue) are shown. B , C . Plots of 53BP1 ( B ) and BRCA1 ( C ) foci per nucleus at 2 h after 6 Gy in DAPI Low and DAPI High cells. D . Cells were treated as in A but fixed after 24 h. E, F . Plots of 53BP1 ( E ) and BRCA1 ( F ) foci per nucleus at 24 h after 6 Gy in DAPI Low and DAPI High cells. For images, scale bar = 20 μm; inset is mean ± SEM foci per nucleus. For plots, red bars indicate mean ± SEM; ****, P < 0.0001; ***, P < 0.001; **, P < 0.01; *, P < 0.05; ns, P > 0.05 (unpaired t-test).

Article Snippet: The primary antibodies used were 53BP1, BRCA1 (Santa Cruz Biotechnology; sc-6954; 1:1,000), and RAD51 (Novus Biologicals; NB100–148; 1:1,000).

Techniques: Irradiation, Staining

Journal: Journal of cell science

Article Title: Cancer metabolism in radiation sensitization: complementary roles of O-GlcNAc transferase (OGT) and PARP1

doi: 10.1242/jcs.264322

Figure Lengend Snippet: A . MCF7 cells were incubated with 10 μM BrdU for 24 h, then treated for 1 h before 6 Gy with DMSO ± 10 μM veliparib or 50 μM PUGNAc + 10 μM veliparib, fixed after 24 h and single strand DNA detected with anti-BrdU antibody under non-denaturing conditions. Representative images show BrdU foci in red with DAPI in blue. B-D . Cells were treated for 1 h before 6 Gy with DMSO ± 10 μM veliparib or 50 μM PUGNAc + 10 μM veliparib, fixed after 24 h and probed for RPA ( B ), RAD51 ( C ) or BRCA1 ( D ). E-H . Plots of BrdU ( E ), RPA ( F ), RAD51 ( G ), and BRCA1 ( H ) foci per nucleus in at 24 h after 6 Gy. For images, scale bar = 20 μm; inset is mean ± SEM foci per nucleus. For plots, red bars indicate mean ± SEM; ****, P < 0.0001; ***, P < 0.001; **, P < 0.01; *, P < 0.05; ns, P > 0.05 (unpaired t-test).

Article Snippet: The primary antibodies used were 53BP1, BRCA1 (Santa Cruz Biotechnology; sc-6954; 1:1,000), and RAD51 (Novus Biologicals; NB100–148; 1:1,000).

Techniques: Incubation

Journal: Journal of cell science

Article Title: Cancer metabolism in radiation sensitization: complementary roles of O-GlcNAc transferase (OGT) and PARP1

doi: 10.1242/jcs.264322

Figure Lengend Snippet: A, B . shScr, shOGT, and shOGA were induced for 48 h with 1 μg/ml doxycycline and cells were treated with 0 or 10 μM veliparib for 1 h prior to 6 Gy, fixed after 24 h and stained for BRCA1. Shown are representative images ( A ) and plots of BRCA1 foci per nucleus ( B ). C, D . Cells were treated as in A and stained for RPA. Shown are representative images ( C ) and plots of RPA foci per nucleus ( D ). E . shScr, shOGT, and shOGA cells were treated as in A and then lightly permeabilized, fixed and then stained with anti-DNA to detect cytosolic DNA (cytoDNA). Shown are representative pseudo-colored images, with cytoDNA in red, DAPI counterstain in blue, and overlays. For images, scale bar = 20 μm; inset is mean ± SEM foci per nucleus. For plots, red bars indicate mean ± SEM; ****, P < 0.0001; ***, P < 0.001; **, P < 0.01; *, P < 0.05; ns, P > 0.05 (unpaired t-test).

Article Snippet: The primary antibodies used were 53BP1, BRCA1 (Santa Cruz Biotechnology; sc-6954; 1:1,000), and RAD51 (Novus Biologicals; NB100–148; 1:1,000).

Techniques: Staining