pp2a inhibition  (Proteostasis Therapeutics)

Journal: Nature Communications

Article Title: The CIP2A-TOPBP1 complex safeguards chromosomal stability during mitosis

doi: 10.1038/s41467-022-31865-5

Figure Lengend Snippet: a Western blots of total cell extract of RPE-1 wild type cells (WT) and RPE-1 CIP2A knock-out cells cells (ΔCIP2A) either Mock treated (-) or treated with the PP2A inhibitor LB-100. b GFP pull-down from 293FT cells transfected with GFP-tagged full-length MDC1 and either control siRNA (siCtrl) or siRNA against CIP2A (siCIP2A). All scale bars = 10 µm. Source data are provided as a Source Data file.

Article Snippet: For cell synchronization in pro-metaphase, cells were incubated with 5 μg/ml Nocodazole (Sigma-Aldrich, M1404) for 16 h. For CRM1 inhibition, 10 μM Leptomycin B (Apollo Scientific) or 10 μM Selinexor (Focus Biomolecules) were added to the medium for 3 h. For PP2A inhibition, 10 nM LB-100 (Selleckchem, S7537) was added for 2 h. mCherry-Fok1 DSB-reporter was induced by treatment with 1 μM Shield-1 and 1 μM 4-hydroxytamoxifen (4-OHT) for 4 h. Irradiation of cells was done in a YXLON, Y.SMART583 custom-made X-ray device or a Faxitron 43855D irradiator.

Techniques: Western Blot, Knock-Out, Transfection, Control

Journal: bioRxiv

Article Title: A cell cycle kinase-phosphatase module restrains PI3K-Akt activity in an mTORC1-dependent manner

doi: 10.1101/2020.11.26.399915

Figure Lengend Snippet: a) Co-depletion of ENSA and ARPP19 proteins in the MDA-MB-231 cell line using specific siRNAs or control siRNAs against luciferase ( Luc ). 72 h after transfection cells were starved for glucose for 1 h and re-stimulated with glucose for 15 min. Total cell extracts were collected and analyzed for the indicated antibodies. b) isgMASTL cells were previously infected with lentiviral supernatants expressing FLAG- and HA-tagged versions of ENSA and ARPP19 phospho-mimetic mutants (S67D and S62D) or GFP as a control. Cells were treated with Dox to induce MASTL CRISPR/Cas9-dependent knock-out, and 72 h later cells were treated as specified in a) . c) Control (Dox −) and MASTL knockout (Dox +) cells were treated with 5 μM fostriecin, 50 nM okadaic acid (OA) or vehicle (Veh) 10 min before insulin stimulation for 30 min. Total lysates were analyzed for the indicated antibodies. Histograms in b, c, represents the quantification of phospho-AKT T308 from 3 independent experiments. Data are mean ± s.e.m. * P <0.05; paired Student’s t -test.

Article Snippet: For PP2A inhibition okadaic acid (Calbiochem) and fostriecin (Santa Cruz Biotechnologies) were used at 50 nM and 5 μM, respectively.

Techniques: Control, Luciferase, Transfection, Infection, Expressing, CRISPR, Knock-Out

Journal: Marine Drugs

Article Title: Evaluation of Rapid, Early Warning Approaches to Track Shellfish Toxins Associated with Dinophysis and Alexandrium Blooms

doi: 10.3390/md16010028

Figure Lengend Snippet: Recovery of certified reference materials from National Research Council of Canada used in multiple analyses to validate protein phosphate inhibition (PP2A) assay kit purchased from Abraxis. CRM, certified reference material; mus-c, contaminated mussel.

Article Snippet: The use of the Abraxis protein phosphate inhibition (PP2A) assay was validated using certified reference materials purchased from the National Research Council of Canada, with reference materials including a naturally-contaminated mussel sample (CRM-DSP-mus-c) and a methanolic calibration standard, yielding recoveries between 100% and 113% and a certified non-toxic mussel homogenate (CRM-zero-mus) yielding concentrations below methodological detection limits ( ).

Techniques: Inhibition

Journal: Marine Drugs

Article Title: Evaluation of Rapid, Early Warning Approaches to Track Shellfish Toxins Associated with Dinophysis and Alexandrium Blooms

doi: 10.3390/md16010028

Figure Lengend Snippet: Total DSP toxins (ng g −1 ) in wild shellfish harvested from regions of New York. Comparison of two extraction methods (double exhaustive and single dispersive) and comparison of two analytical methods (protein phosphatase inhibition assay, PP2A; and liquid chromatography mass spectrometry, LC/MS). Bold indicates over the federal closure limit (160 ng g −1 ).

Article Snippet: The use of the Abraxis protein phosphate inhibition (PP2A) assay was validated using certified reference materials purchased from the National Research Council of Canada, with reference materials including a naturally-contaminated mussel sample (CRM-DSP-mus-c) and a methanolic calibration standard, yielding recoveries between 100% and 113% and a certified non-toxic mussel homogenate (CRM-zero-mus) yielding concentrations below methodological detection limits ( ).

Techniques: Comparison, Extraction, Inhibition, Liquid Chromatography, Mass Spectrometry

Journal: Marine Drugs

Article Title: Evaluation of Rapid, Early Warning Approaches to Track Shellfish Toxins Associated with Dinophysis and Alexandrium Blooms

doi: 10.3390/md16010028

Figure Lengend Snippet: Total DSP toxins (ng g −1 ) in ( A ) softshell clams ( n = 9) and ( B ) various shellfish species ( n = 6) from New York embayments analyzed via LCMS and PP2A. Each point represents >6 pooled individuals.

Article Snippet: The use of the Abraxis protein phosphate inhibition (PP2A) assay was validated using certified reference materials purchased from the National Research Council of Canada, with reference materials including a naturally-contaminated mussel sample (CRM-DSP-mus-c) and a methanolic calibration standard, yielding recoveries between 100% and 113% and a certified non-toxic mussel homogenate (CRM-zero-mus) yielding concentrations below methodological detection limits ( ).

Techniques: