Journal: bioRxiv
Article Title: GTP signaling links metabolism, DNA repair, and responses to genotoxic stress
doi: 10.1101/2023.04.12.536297
Figure Lengend Snippet: (A&B) Cells were treated with the phosphatase inhibitors okadaic acid (OA; 15 nm) or fostricein (Fos; 100 nm) 1 h before RT (A), or first transfected with pan PP1 siRNA, mixture of PP2A catalytic (PP2A-C) subunit α/β, PP4 and PP5 for 48 h and irradiated after transfection (B), followed by cell harvesting 4 h post-RT for immunoblot. (C-F) Cells were transfected with constitutively active Rac1-Q61L and then treated with okadaic acid (15 nm) or fostricein (100 nm) 1 h before RT (C&D), or transfected with individual phosphatase siRNA pool, along with overexpression of Rac1-Q61L or control (E&F), followed by immunoblot (C&E) or γ-H2AX foci IF staining (D&F). Data are presented as mean ± SEM from three biologically independent experiments for Figure D, F and Figure A, B, C, E are representative figures from three biologically independent experiments. Two-tailed unpaired student’s t test **p < 0.01, ***p < 0.001, ****p < 0.0001.
Article Snippet: SiRNAs of pan PP1 (a pool of 6 siRNAs, sc-43545), PP2A-Cα (a pool of 3 siRNAs, sc-43509), PP2A-Cβ (a pool of 3 siRNAs, sc-36301), PP4 (a pool of 3 siRNAs, sc-39202), PP5 (a pool of 3 siRNAs, sc-44602), Abi-1 (sc-417534-NIC) double nickase CRISPR/CAS9 knockout plasmids and control (sc-437281) were obtained from Santa Cruz Biotechnology.
Techniques: Transfection, Irradiation, Cell Harvesting, Western Blot, Over Expression, Control, Staining, Two Tailed Test
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