Journal: bioRxiv

Article Title: Imaging-Guided Metabolic Radiosensitization of Pediatric Rhabdoid Tumors

doi: 10.1101/2024.08.09.607364

Figure Lengend Snippet: (A) Whole tissue imaging and ( B ) confocal micrographs of γ-H2AX immunofluorescence staining of all treatment groups on Day 9. (Insets) Stained A-204 tissue at high magnification. Scale bar = 50 µm for confocal images and scale bar = 1000 µm for whole tumor sections. (C) Quantification of γ-H2AX staining in whole tumor sections ( a ) and confocal micrographs ( b ) of all treatment groups. (D) H&E staining for heart, liver, kidney and muscle from various treatment groups. Scale bar=100 µm. Data represented as mean ± SD.

Article Snippet: Alexa Fluor 488 AffiniPure Donkey Anti-Rat lgG(H+L) (Lot 163136) and Rhodamine (TRITC) AffiniPure Donkey Anti-Rat lgG(H+L) (Lot 157518) were obtained from Jackson ImmunoResearch Inc. Histone H2AX antibody (NB100-384) was purchased from Novus Biologicals and secondary antibody Donkey anti-Rabbit lgG (H+L) Dylight594 (SA5-10040) from Invitrogen.

Techniques: Imaging, Immunofluorescence, Staining

Journal: European Journal of Nuclear Medicine and Molecular Imaging

Article Title: Combining [ 177 Lu]Lu-DOTA-TOC PRRT with PARP inhibitors to enhance treatment efficacy in small cell lung cancer

doi: 10.1007/s00259-024-06844-1

Figure Lengend Snippet: Determination of γH2AX foci formation and clonogenic survival in H446 cells. (A) Exemplary images of H446 cells after mono- or combination therapy directly after treatment or after a 23 h post-incubation time. (B) Number of γH2AX foci per cell after mono- or combination therapy directly after treatment or after a 23 h post-incubation time in medium. Displayed are mean ± SEM from 3 individual experiments. (C) Surviving fractions of H446 cells mono- or combination therapy. Displayed are mean values from 2 independent experiments

Article Snippet: 1:150 dilution), γH2AX (pSER139, Novus #NB100-2280, 1:500 dilution) with optimized protocols of the Comparative Experimental Pathology Core Facility.

Techniques: Incubation

Journal: European Journal of Nuclear Medicine and Molecular Imaging

Article Title: Combining [ 177 Lu]Lu-DOTA-TOC PRRT with PARP inhibitors to enhance treatment efficacy in small cell lung cancer

doi: 10.1007/s00259-024-06844-1

Figure Lengend Snippet: Immunohistochemistry of H69 tumors. Exemplary stainings of H69 tumors obtained from the single dose PRRT study ( n = 1 animal/group) for SSTR2, PARP1, γH2AX (DNA damage marker) and cleaved caspase 3 (apoptosis marker) are displayed. Scale bar = 100 µm. Brown color indicates positivity for the respective marker

Article Snippet: 1:150 dilution), γH2AX (pSER139, Novus #NB100-2280, 1:500 dilution) with optimized protocols of the Comparative Experimental Pathology Core Facility.

Techniques: Immunohistochemistry, Marker

Journal: Cancer Research

Article Title: Stalled Replication Induces p53 Accumulation through Distinct Mechanisms from DNA Damage Checkpoint Pathways

doi: 10.1158/0008-5472.can-05-1790

Figure Lengend Snippet: Figure 3. Induction of p53 by hydroxyurea correlates with the activation of MRN-ATM pathway but is insensitive to caffeine. A, ATM is autophosphorylated after DNA damage and replication block. Normal lymphoblastoid cells (WT) were treated with the indicated agents for 24 hours. The levels of phospho-ATMSer1981 and total ATM were detected by immunoblotting. Extracts of ataxia telangiectasia lymphoblastoid cells act as negative controls. B, hydroxyurea induces significantly fewer DNA breakage than Adriamycin. Wild-type lymphoblastoid cells were mock treated or treated with hydroxyurea or Adriamycin for 16 hours. The cells were then harvested for single-cell gel electrophoresis assay. Representational images are shown and tail movements were quantified. Columns, mean of three independent experiments; bars, SD. C, hydroxyurea-induced ATM, p53, and histone H2AX phosphorylation are not suppressed by caffeine. Wild-type lymphoblastoid cells were treated with buffer or hydroxyurea for 6 hours before buffer or caffeine was added for another 6 hours. Cell extracts were prepared and the expression of p53, phospho-p53Ser20, phospho-ATMSer1981, ATM, phospho-histone H3Ser10, and phospho-histone H2AXSer139 was detected by immunoblotting. Uniform loading of lysates was confirmed by immunoblotting for CDC2.

Article Snippet: Polyclonal antibodies against Ser139-phosphorylated histone H2AX were either gifts from Junjie Chen (Mayo Clinic, Rochester, MN) or obtained from Bethyl Laboratories (Montgomery, TX).

Techniques: Activation Assay, Blocking Assay, Western Blot, Single Cell Gel Electrophoresis, Phospho-proteomics, Expressing