Journal: Cell reports

Article Title: PPARγ Interaction with UBR5/ATMIN Promotes DNA Repairto Maintain Endothelial Homeostasis

doi: 10.1016/j.celrep.2019.01.013

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rat Anti-ORC2 Monoclonal Antibody, Unconjugated, Clone 3G6 , Cell Signaling Technology , Cat#4736, RRID:AB_2157716.

Techniques: Control, Recombinant, In Vitro, Transfection, Flow Cytometry, Single Cell Gel Electrophoresis, Mass Spectrometry, Sequencing, Real-time Polymerase Chain Reaction, Cloning, Plasmid Preparation, Software

Journal: Journal of Biological Chemistry

Article Title: Control of DNA Replication by the Nucleus/Cytoplasm Ratio in Xenopus

doi: 10.1074/jbc.m113.499012

Figure Lengend Snippet: FIGURE 7. High N/C ratio nuclei contain lower amounts of PP2A. A and B, nuclei were isolated from extracts containing either 1,000 or 10,000 nuclei/l and analyzed by immunoblot for PP2A components and ORC2. For 10,000 nuclei/l, two different isolations were performed. In the first, an equal number of nuclei were isolated relative to the 1,000 nuclei/l sample. In the second, nuclei were isolated from an equal volume of extract relative to the 1,000 nuclei/l sample. A sample containing no nuclei is included to control for residual cytoplasmic protein. The results from two independent experiments are shown. C, after the removal of nuclei, cytoplasm from the samples in B was analyzed for PP2A C content. D, extracts were prepared with either 1,000 or 10,000 nuclei/l and supplemented with either control buffer or 478 nM rPP2A C (Xenopus). An equal number of nuclei were isolated from each condition and blotted for the indicated proteins.

Article Snippet: Chk1 (FL-476) and PP2A A (6G3) antibodies were purchased from Santa Cruz Biotechnology, Inc. Antibodies recognizing ORC2 were affinity-purified from rabbit antiserum raised against Xenopus ORC2, which was produced in bacteria.

Techniques: Isolation, Western Blot, Control

Journal: Journal of Biological Chemistry

Article Title: Control of DNA Replication by the Nucleus/Cytoplasm Ratio in Xenopus

doi: 10.1074/jbc.m113.499012

Figure Lengend Snippet: FIGURE 8. B55 levels control the amount of DNA synthesis at high N/C ratio. A, interphase extracts were immunodepleted of B55 or mock-depleted. Sperm chromatin was added to 6,000/l, and DNA synthesis was measured. The data shown are from a single experiment and are representative of multiple, independent experiments. B, the extent of depletion was monitored by immunoblot. The asterisk marks an unknown nonspecific band. C, mock-depleted or B55-depleted extracts were supplemented with 1,000 nuclei/l. Nuclei were then isolated after 83 min and blotted for PP2A C and ORC2. D and E, cytostatic factor-arrested extracts were incubated with up to 100 or 133 ng/l B55 mRNA for 2 h at room temperature, activated with CaCl2, and then supplemented with sperm chromatin to 10,000/l. DNA synthesis was measured as indicated. A sample containing only 1,000 nuclei/l is shown for comparison. The results from two independent experiments are shown. F, production of B55 was monitored by immunoblot. The asterisk marks an unknown nonspecific band. G, the B55 bands in F were quantified, and the relative intensities of each are shown. a.u., absorbance units.

Article Snippet: Chk1 (FL-476) and PP2A A (6G3) antibodies were purchased from Santa Cruz Biotechnology, Inc. Antibodies recognizing ORC2 were affinity-purified from rabbit antiserum raised against Xenopus ORC2, which was produced in bacteria.

Techniques: Control, DNA Synthesis, Western Blot, Isolation, Incubation, Comparison