anti brca1  (Santa Cruz Biotechnology)

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

Journal: Nature Communications

Article Title: Combination of PARP and KRAS G12D inhibitors enhances therapeutic efficacy by exploiting vulnerabilities in PDAC

doi: 10.1038/s41467-026-69695-4

Figure Lengend Snippet: a – d Western blot analysis of BRCA1, RPA32, RAD51, total ERK, total AKT, phospho-ERK, and phospho-AKT in AsPC-1 ( a ), PANC-1 ( b ), SW1990 ( c ), and KPC1199 ( d ) cells treated with vehicle (DMSO) or MRTX1133 at indicated concentrations for 24 h. GAPDH or β-actin served as the loading control. Similar results were obtained from three independent experiments. e – h Immunofluorescence assay for DNA damage marker in KRAS G12D PDAC cell lines. Representative micrographs ( e ) and quantification data for MDC1, FK2, BRCA1, RPA32, and RAD51 foci formation ( f – h ) in the indicated cell lines with and without MRTX1133 treatment (50 nM). Quantification shows the number of foci per cell. Data are presented as Mean ± SEM of n = 3 biologically independent experiments. Statistical significance was determined by an unpaired, two-tailed Student’s t -test. The scale bar indicates 10 µm. i Schematic of the DR-GFP reporter system. j HR efficiency was measured in AsPC-1, PANC-1, and SW1990 cells using the DR-GFP reporter assay after treatment with MRTX1133 (10 nM, 50 nM) or vehicle. Data are presented as mean ± SEM of n = 3 biologically independent experiments. Statistical significance compared to the vehicle control was determined by one-way ANOVA. Source data are provided as a Source Data file.

Article Snippet: Antibodies were obtained from the following sources: γ-H2AX (Millipore, 05-636), MDC1 (Millipore, 05-1572), FK2 (Millipore, 04-263), BRCA1 (Santa Cruz Biotechnology, sc-6954), RPA32 (Santa Cruz, sc-56770) and RAD51 (GeneTex, GTX100469).

Techniques: Western Blot, Control, Immunofluorescence, Marker, Two Tailed Test, Reporter Assay

Journal: Nature Communications

Article Title: Combination of PARP and KRAS G12D inhibitors enhances therapeutic efficacy by exploiting vulnerabilities in PDAC

doi: 10.1038/s41467-026-69695-4

Figure Lengend Snippet: a – c Dose-dependent effects of MRTX1133 on HR and signaling proteins. Parental (P) and MRTX1133-resistant (R) variants of AsPC-1, PANC-1, and SW1990 cells were treated with the indicated concentrations of MRTX1133 for 24 h. Data are representative of three independent experiments with similar results. d – f Time-dependent effects of MRTX1133 on HR and signaling proteins. Parental (P) and resistant (R) cells were treated with 50 nM MRTX1133 for the indicated times (6–72 h). Protein levels were assessed by Western blot as described above. Data are representative of three independent experiments with similar results. g , h Immunofluorescence assay for HR-related protein in MRTX1133-resistant KRAS G12D PDAC cell lines. Representative immunofluorescence images ( g ) and quantification ( h ) of BRCA1, RPA32, and RAD51 foci in resistant cell lines are shown. Data are presented as mean ± SEM of n = 3 biologically independent experiments. Statistical significance was determined by a two-sided unpaired Student’s t -test. i MRTX1133 treatment reduces HR repair efficiency. Parental and resistant cells were treated with MRTX1133 (50 nM) for 24 h, and HR efficiency was measured using a DR-GFP reporter assay. Data are presented as mean ± SEM of n = 3 biologically independent experiments. Statistical significance was determined by the unpaired, two-tailed Student’s t -test. j – l Synergistic anti-tumor effect of MRTX1133 combined with Olaparib in resistant pancreatic cancer cells. Three-dimensional synergy plots show the interaction between MRTX1133 and Olaparib in AsPC-1-R ( j ), PANC-1-R ( k ), and SW1990-R ( l ) cells after 144 h of treatment. Red peaks indicate synergy, and green valleys indicate antagonism. Data represent the mean of three independent biological replicates ( n = 3). Statistical significance of the synergy score was determined by a one-sample t -test comparing the mean synergy score against a reference value of 0 (two-sided). Source data are provided as a Source Data file.

Article Snippet: Antibodies were obtained from the following sources: γ-H2AX (Millipore, 05-636), MDC1 (Millipore, 05-1572), FK2 (Millipore, 04-263), BRCA1 (Santa Cruz Biotechnology, sc-6954), RPA32 (Santa Cruz, sc-56770) and RAD51 (GeneTex, GTX100469).

Techniques: Western Blot, Immunofluorescence, Reporter Assay, Two Tailed Test