Journal: Cell reports

Article Title: Heterogeneity and transcriptional drivers of triple-negative breast cancer

doi: 10.1016/j.celrep.2023.113564

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Mouse monoclonal anti-cPARP (164Dy) , BD Biosciences , Cat# 552596; RRID: AB_394437.

Techniques: Recombinant, Mass Spectrometry, shRNA, Plasmid Preparation

Journal: Interactive Cardiovascular and Thoracic Surgery

Article Title: Povidone-iodine results in rapid killing of thymic epithelial tumour cells through cellular fixation ^†

doi: 10.1093/icvts/ivy248

Figure Lengend Snippet: Povidone-iodine results in rapid killing of thymic epithelial tumour cells through cellular fixation. (A) Phase-contrast microscopic findings. Under low concentration of PVP-I, formazan crystals (purple) were visible in MeT-5A, IU-TAB-1 and Ty-82 cells evaluated in an MTT assay. Following the addition of DMSO to the MTT assay for purposeful destruction of the lipid bilayer structure of the cell membrane 4 h after incubation of cells in MTT solution, microscopy demonstrated that the cell structure was intact 5 min following 1% PVP-I treatment of thymic epithelial tumour cells. (B) Intracellular staining of cPARP for both IU-TAB-1 and Ty-82 after 1% PVP-I treatment showed to resemble cPARP staining following treatment with known fixatives, 4% paraformaldehyde (PFA) and 75% ethyl alcohol (EtOH). cPARP: cleaved poly-ADP-ribose polymerase; DMSO: dimethylsulphoxide; EtOH: ethyl alcohol; PFA: paraformaldehyde; PVP-I: povidone-iodine.

Article Snippet: Intracellular cleaved poly-ADP-ribose polymerase (cPARP) staining was evaluated using anti-human cPARP (Cat#. 558710, BD biosciences).

Techniques: Concentration Assay, MTT Assay, Incubation, Microscopy, Staining

Journal: Cell Death and Differentiation

Article Title: PCAF ubiquitin ligase activity inhibits Hedgehog/Gli1 signaling in p53-dependent response to genotoxic stress

doi: 10.1038/cdd.2013.120

Figure Lengend Snippet: PCAF prevents the mitogenic and prosurvival activity of Gli1 in response to genotoxic stress. (a) Protein levels of Gli1 and PCAF in Mb tumor samples from Ptch1+/− or Ptch1+/−;p53−/− mouse model. (b) D283 medulloblastoma cells were transfected for 24 h with Gli1 or control vector and then treated with 2 μM doxorubicin. Percent of BrdU incorporation was assessed in the population of transfected cells to monitor cell proliferation. *P<0.05, doxo versus control; **P<0.05 Gli1+doxo versus empty+doxo. (c) D283 cells were transfected and treated as in (b). Apoptosis was assessed in the population of transfected cells by evaluation of cPARP and nuclear morphology. *P<0.05, doxo versus control; **P<0.05, Gli1+doxo versus empty+doxo. (d) D283 medulloblastoma cells were transfected with siPCAF or siCTR and then treated with 2 μM doxorubicin. Percent of BrdU incorporation was assessed as described in (b). *P<0.05, doxo versus control; **P<0.05 siPCAF+doxo versus siCTR+doxo. (e) D283 cells were transfected and treated as in (d). Apoptosis was evaluated as described in (c). *P<0.05, doxo versus control; **P<0.05 siPCAF+doxo vs siCTR+doxo. (f) Daoy medulloblastoma cells were transfected with the specified siRNA and with the indicated plasmids. Percent of BrdU incorporation was assayed to monitor cell proliferation. *P<0.05, siGli1 versus siCTR or PCAF/siCTR versus empty/siCTR. (g) Daoy cells were transfected with the indicated plasmids, grown with G418 for 2 weeks, fixed and stained with crystal violet. Colony formation assay was performed to monitor cell proliferation. Counts from the colony formation assays and representative images from experiments are shown. *P<0.05, PCAF or Gli1 versus empty. (h) Cells derived from triplicate plates from the colony assays were grown in culture as monolayers, and cell growth rates were determined by analyzing cumulative cell number over time. Cells were counted in triplicate. Unless otherwise indicated, all experiments were performed in triplicate, and mean±S.D. is shown

Article Snippet: Apoptosis was measured by analysis of nuclear morphology and immunofluorescence using rabbit anti-cPARP antibody (Promega).

Techniques: Activity Assay, Transfection, Control, Plasmid Preparation, BrdU Incorporation Assay, Staining, Colony Assay, Derivative Assay