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Image Search Results
Journal: The Journal of Cell Biology
Article Title: A PP2A-B55 recognition signal controls substrate dephosphorylation kinetics during mitotic exit
doi: 10.1083/jcb.201606033
Figure Lengend Snippet: B55 regulates the timing of nuclear pore reassembly and nuclear transport during mitotic exit. (A) Nuclear pore proteins identified as candidate B55 substrates. PPSIM plots for NUP153 pS257 and pS240 are shown; the dotted line indicates the profile for PRC1 pT470 and TPX2 pT369. PPSIM kinetic profiles for TMPO pS362, LEMD2 pS166, NDC1 pT414 and NUP107 pT46 are displayed on the right. Error bars indicate the SEM (siControl, n = 5; siB55, n = 3; siMASTL, n = 2). (B) The localization of NUP153 (red) and FG-repeat nucleoporins (mAb414; green) is shown in anaphase and telophase control cells (siControl) and cells depleted of Greatwall (siMASTL) or all B55 regulatory subunits (siB55 all). DNA was stained with DAPI (blue). (C) Reformation of nuclear pores and the resumption of nuclear import were followed by live-cell imaging of cells expressing GFP-NUP153 and the mCherry-Importin β binding domain (IBB), respectively. Images are shown every 3 min from the onset of anaphase ( t = 0 min) until 21 min for control cells (siControl) and cells depleted of Greatwall (siMASTL) or all B55 regulatory subunits (siB55 all). (D and E) Nuclear pore reassembly was measured using the GFP-NUP153 signal surrounding the segregating chromatin (D), and nuclear import was defined as the mCherry-IBB signal within this region (E). Error bars indicate SEM ( n = 6).
Article Snippet: The commercial antibodies used were PRC1 pT481 (2189–1; Epitomics), cyclin B1 (05–373; EMD Millipore), tubulin (T6199; Sigma-Aldrich), PPP2CA (610555; BD), PPP2R1A (sc-6112; Santa Cruz Biotechnology, Inc.), PPP2R2A (5689S; Cell Signaling Technology), PPP2R5A (A300-967A; Bethyl Laboratories, Inc.), PPP2R5C, (sc-374380; Santa Cruz Biotechnology, Inc.), PPP2R5D (A301-100A; Bethyl Laboratories, Inc.), MASTL (A302-190A; Bethyl Laboratories, Inc.; or ab135637; Abcam), nucleoporin FG-repeat mAb414 (ab24609; Abcam), lamin B receptor (1398–1; Epitomics), AHCTF1 (A300-166A; Bethyl Laboratories, Inc.),
Techniques: Control, Staining, Live Cell Imaging, Expressing, Binding Assay
Journal: The Journal of Cell Biology
Article Title: A PP2A-B55 recognition signal controls substrate dephosphorylation kinetics during mitotic exit
doi: 10.1083/jcb.201606033
Figure Lengend Snippet: A BPR in NUP153 promotes timely activation in anaphase. (A) The localization of wild-type, (B) S257A, and (C) BPR mutant (R 214 /K 227 /K 228 were mutated to alanine) GFP-NUP153 at different stages of mitosis is shown compared with FG-repeat nucleoporins (mAb414), RanBP2, and DNA. (D) HeLa cells expressing GFP-NUP153 wild-type, S257A, and BPR mutants were imaged every minute through mitosis. Images of G2 cells before mitotic entry, and at the times indicated from the onset of anaphase, are shown. (E) Nuclear pore reassembly was measured using the GFP-NUP153 signal surrounding the segregating chromatin. Error bars indicate SEM ( n = 6).
Article Snippet: The commercial antibodies used were PRC1 pT481 (2189–1; Epitomics), cyclin B1 (05–373; EMD Millipore), tubulin (T6199; Sigma-Aldrich), PPP2CA (610555; BD), PPP2R1A (sc-6112; Santa Cruz Biotechnology, Inc.), PPP2R2A (5689S; Cell Signaling Technology), PPP2R5A (A300-967A; Bethyl Laboratories, Inc.), PPP2R5C, (sc-374380; Santa Cruz Biotechnology, Inc.), PPP2R5D (A301-100A; Bethyl Laboratories, Inc.), MASTL (A302-190A; Bethyl Laboratories, Inc.; or ab135637; Abcam), nucleoporin FG-repeat mAb414 (ab24609; Abcam), lamin B receptor (1398–1; Epitomics), AHCTF1 (A300-166A; Bethyl Laboratories, Inc.),
Techniques: Activation Assay, Mutagenesis, Expressing