Journal: Molecular oncology

Article Title: Palbociclib induces activation of AMPK and inhibits hepatocellular carcinoma in a CDK4/6-independent manner.

doi: 10.1002/1878-0261.12072

Figure Lengend Snippet: Fig. 3. PP5 mediates palbociclib-induced AMPK activation, autophagy, and apoptosis. (A) PP5 overexpression suppresses the effect of palbociclib on pAMPKa, autophagy, and apoptosis. Hep3B cells were transfected with vector or DDK-PP5 for 24 h and treated with palbociclib (15 lM) for another 24 h. (B) Pretreatment with arachidonic acid (AA), a PP5 activator, reversed the effect of palbociclib on pAMPKa, autophagy, and apoptosis. Hep3B cells were pretreated with AA (100 lM) for 4 h and then treated with palbociclib for 24 h. Autophagy was determined by LC3 immunoblotting. Apoptotic cells were measured by flow cytometry. *P < 0.05. (C) PP5 activity in Hep3B and PLC5 cells. (D) Palbociclib inhibits PP5 activity in PP5-containing Hep3B lysate. (E) Palbociclib suppresses activity of GST-PP5. Cantharidin, a known Ser/Thr phosphatase inhibitor, served as a positive control. *P < 0.05

Article Snippet: PP5 antibody (C-20) (sc-32588) was used for immunoblotting, and the PP5 antibody (H-7) (sc271816) used for immunoprecipitation was obtained from Santa Cruz Biotechnology (San Diego, CA, USA).

Techniques: Activation Assay, Over Expression, Transfection, Plasmid Preparation, Western Blot, Cytometry, Activity Assay, Positive Control

Journal: Molecular oncology

Article Title: Palbociclib induces activation of AMPK and inhibits hepatocellular carcinoma in a CDK4/6-independent manner.

doi: 10.1002/1878-0261.12072

Figure Lengend Snippet: Fig. 5. In vivo effects of palbociclib in Huh7 xenograft nude mice. (A,B) Palbociclib suppresses Huh7 xenograft tumor growth and weight. Nude mice were subcutaneously implanted with 5 9 106 Huh7 cells. The mice received vehicle or palbociclib (150 mgkg1) orally every three days. The tumor area was measured twice a week. At the end of treatment, the tumors were harvested and the tumor weights were determined before lysis. Points/bars, mean (n = 5); bars, SD. **P < 0.01. (C) Western blot analysis of pAMPK, AMPK, and LC3 in Huh7 tumors. (D) PP5 activity in Huh7 tumors. *P < 0.05.

Article Snippet: PP5 antibody (C-20) (sc-32588) was used for immunoblotting, and the PP5 antibody (H-7) (sc271816) used for immunoprecipitation was obtained from Santa Cruz Biotechnology (San Diego, CA, USA).

Techniques: In Vivo, Lysis, Western Blot, Activity Assay

Journal: Molecular oncology

Article Title: Palbociclib induces activation of AMPK and inhibits hepatocellular carcinoma in a CDK4/6-independent manner.

doi: 10.1002/1878-0261.12072

Figure Lengend Snippet: Fig. 6. Expression of PP5 in human HCC tissues. (A) Expression of PP5 in HCC tumors and normal liver tissue. The protein expressions of PP5 in the HCC tumors and its adjacent normal part were analyzed in 153 patients with HCC by IHC. Representative images were shown here. (B) PP5 expression in HCC tumors is significantly higher in HCC tumors than in normal liver tissue. The expressions of PP5 were quantified by H-score and compared by Student’s t-test. P < 0.001. Bar, mean; error bar, SE. ***P < 0.001. (C) PP5 expression in paired tumor and normal liver tissue. Each dot represented the H-score obtained from indicated tissues, and each line linked an individual patient.

Article Snippet: PP5 antibody (C-20) (sc-32588) was used for immunoblotting, and the PP5 antibody (H-7) (sc271816) used for immunoprecipitation was obtained from Santa Cruz Biotechnology (San Diego, CA, USA).

Techniques: Expressing

Journal: Marine Drugs

Article Title: Structure–Activity Relationship Studies Using Natural and Synthetic Okadaic Acid/Dinophysistoxin Toxins

doi: 10.3390/md14110207

Figure Lengend Snippet: Effect of various DSP toxins on protein phosphatase activity. The activities of protein phosphatases ( A ) PP2a, ( B ) PP1, and ( C ) PP5 were assessed in the presence of various concentrations of natural OA, natural DTX1, natural DTX2, synthetic DTX2, or synthetic 2- epi -DTX2. All data (mean ± SE; n = 3 or 4) were normalized to the control. Three-parameter, variable slope, non-linear dose-response analysis was performed and IC 50 and 95% confidence interval values were calculated . Note: Some of these data have been published previously in Pang et al. .

Article Snippet: Briefly, the assay tests the ability of toxins to inhibit the activity of purified PP2A (EMD Millipore Corporation, Darmstadt, Germany, cat. #14-111; human red blood cells), purified PP1 (EMD Millipore Corporation, Darmstadt, Germany, cat. #14-110; rabbit skeletal muscle), or purified PP5 (EMD Millipore Corporation, Darmstadt, Germany, cat. #14-778; recombinant enzyme expressed in E. coli cells) against the fluorimetric substrate, 6,8-difluro-4-methyl umbelliferyl phosphate (DiFMUP; 50 μM final) (Invitrogen ThermoFisher Scientific, Waltham, MA, USA, cat. #D6567).

Techniques: Activity Assay, Control

Journal: Marine Drugs

Article Title: Structure–Activity Relationship Studies Using Natural and Synthetic Okadaic Acid/Dinophysistoxin Toxins

doi: 10.3390/md14110207

Figure Lengend Snippet: Calculated IC 50 values and relative potencies for natural OA, DTX1, and DTX2, and synthetic DTX2, and 2 -epi -DTX2, based on PP2a, PP1, and PP5 inhibition.

Article Snippet: Briefly, the assay tests the ability of toxins to inhibit the activity of purified PP2A (EMD Millipore Corporation, Darmstadt, Germany, cat. #14-111; human red blood cells), purified PP1 (EMD Millipore Corporation, Darmstadt, Germany, cat. #14-110; rabbit skeletal muscle), or purified PP5 (EMD Millipore Corporation, Darmstadt, Germany, cat. #14-778; recombinant enzyme expressed in E. coli cells) against the fluorimetric substrate, 6,8-difluro-4-methyl umbelliferyl phosphate (DiFMUP; 50 μM final) (Invitrogen ThermoFisher Scientific, Waltham, MA, USA, cat. #D6567).

Techniques: Inhibition

Journal: Cell reports

Article Title: Post-translational Regulation of FNIP1 Creates a Rheostat for the Molecular Chaperone Hsp90

doi: 10.1016/j.celrep.2019.01.018

Figure Lengend Snippet:

Article Snippet: Rabbit anti-PP5 , Cell Signaling Technology , Cat# 2289; RRID:AB_2168757.

Techniques: Virus, Recombinant, Mass Spectrometry, Software