Journal: Pharmaceutics

Article Title: Validation of an LC-MS/MS Method for the Simultaneous Intracellular Quantification of the CDK4/6 Inhibitor Abemaciclib and the EZH2 Inhibitors GSK126 and Tazemetostat

doi: 10.3390/pharmaceutics17040433

Figure Lengend Snippet: Chemical structure, molecular formulae and average molecular weights of abemaciclib, GSK126, tazemetostat and the internal standard palbociclib.

Article Snippet: Abemaciclib and GSK126 were ordered from LKT Laboratories Inc. (St. Paul, MN, USA).

Techniques:

Journal: Pharmaceutics

Article Title: Validation of an LC-MS/MS Method for the Simultaneous Intracellular Quantification of the CDK4/6 Inhibitor Abemaciclib and the EZH2 Inhibitors GSK126 and Tazemetostat

doi: 10.3390/pharmaceutics17040433

Figure Lengend Snippet: Linear binary gradient for the analysis of abemaciclib, GSK126 and tazemetostat (eluent A: water with 0.1% formic acid; eluent B: acetonitrile with 0.1% formic acid).

Article Snippet: Abemaciclib and GSK126 were ordered from LKT Laboratories Inc. (St. Paul, MN, USA).

Techniques:

Journal: Pharmaceutics

Article Title: Validation of an LC-MS/MS Method for the Simultaneous Intracellular Quantification of the CDK4/6 Inhibitor Abemaciclib and the EZH2 Inhibitors GSK126 and Tazemetostat

doi: 10.3390/pharmaceutics17040433

Figure Lengend Snippet: Mass spectrometric parameters for the detection of tazemetostat, abemaciclib, GSK126 and palbociclib.

Article Snippet: Abemaciclib and GSK126 were ordered from LKT Laboratories Inc. (St. Paul, MN, USA).

Techniques:

Journal: Pharmaceutics

Article Title: Validation of an LC-MS/MS Method for the Simultaneous Intracellular Quantification of the CDK4/6 Inhibitor Abemaciclib and the EZH2 Inhibitors GSK126 and Tazemetostat

doi: 10.3390/pharmaceutics17040433

Figure Lengend Snippet: Calibration range and quality control level concentrations of abemaciclib, GSK126 and tazemetostat.

Article Snippet: Abemaciclib and GSK126 were ordered from LKT Laboratories Inc. (St. Paul, MN, USA).

Techniques: Control, Concentration Assay

Journal: Pharmaceutics

Article Title: Validation of an LC-MS/MS Method for the Simultaneous Intracellular Quantification of the CDK4/6 Inhibitor Abemaciclib and the EZH2 Inhibitors GSK126 and Tazemetostat

doi: 10.3390/pharmaceutics17040433

Figure Lengend Snippet: Representative chromatograms of cell suspensions in 20 mM Tris buffer, pH 7.4, spiked with blank methanol ( A ), only palbociclib (IS) ( B ) or palbociclib (IS) and the three analytes (abemaciclib, GSK126, tazemetostat) at lower limit of quantification (LLOQ, ( C )) or upper limit of quantification (ULOQ, ( D )). The mass transition that applies to the respective line of chromatograms is listed on the left.

Article Snippet: Abemaciclib and GSK126 were ordered from LKT Laboratories Inc. (St. Paul, MN, USA).

Techniques:

Journal: Pharmaceutics

Article Title: Validation of an LC-MS/MS Method for the Simultaneous Intracellular Quantification of the CDK4/6 Inhibitor Abemaciclib and the EZH2 Inhibitors GSK126 and Tazemetostat

doi: 10.3390/pharmaceutics17040433

Figure Lengend Snippet: Representative calibration curves of abemaciclib, GSK126 and tazemetostat.

Article Snippet: Abemaciclib and GSK126 were ordered from LKT Laboratories Inc. (St. Paul, MN, USA).

Techniques:

Journal: Pharmaceutics

Article Title: Validation of an LC-MS/MS Method for the Simultaneous Intracellular Quantification of the CDK4/6 Inhibitor Abemaciclib and the EZH2 Inhibitors GSK126 and Tazemetostat

doi: 10.3390/pharmaceutics17040433

Figure Lengend Snippet: Within-run and between-run accuracy and precision.

Article Snippet: Abemaciclib and GSK126 were ordered from LKT Laboratories Inc. (St. Paul, MN, USA).

Techniques: Control

Journal: Pharmaceutics

Article Title: Validation of an LC-MS/MS Method for the Simultaneous Intracellular Quantification of the CDK4/6 Inhibitor Abemaciclib and the EZH2 Inhibitors GSK126 and Tazemetostat

doi: 10.3390/pharmaceutics17040433

Figure Lengend Snippet: Stability of abemaciclib, GSK126 and tazemetostat.

Article Snippet: Abemaciclib and GSK126 were ordered from LKT Laboratories Inc. (St. Paul, MN, USA).

Techniques: Control

Journal: Pharmaceutics

Article Title: Validation of an LC-MS/MS Method for the Simultaneous Intracellular Quantification of the CDK4/6 Inhibitor Abemaciclib and the EZH2 Inhibitors GSK126 and Tazemetostat

doi: 10.3390/pharmaceutics17040433

Figure Lengend Snippet: Intracellular concentrations of abemaciclib (A), GSK126 (G) and tazemetostat (T) in GBM06 and GBM15 cells. Cells were incubated with either 1 µM abemaciclib, 10 µM GSK126, 10 µM tazemetostat or a combination of 1 µM abemaciclib plus 10 µM GSK126 or 10 µM tazemetostat for 72 h, followed by lysis and LC-MS/MS analysis of intracellular drug concentrations. Data are mean + SD of n = 4 independent experiments.

Article Snippet: Abemaciclib and GSK126 were ordered from LKT Laboratories Inc. (St. Paul, MN, USA).

Techniques: Incubation, Lysis, Liquid Chromatography with Mass Spectroscopy

Journal: Heliyon

Article Title: Transforming growth factor- β mediated regulation of epigenome is required for epithelial to mesenchymal transition associated features in liver cancer cells

doi: 10.1016/j.heliyon.2023.e14665

Figure Lengend Snippet: Pharmacological or genetic ablation of EZH2 alters TGF-β induced effects. (a) Protein levels of EZH2 and H3K27me3 post TGF-β or TGF-β with GSK-126 exposure. GSK-126 was added 2 h before TGF-β treatment. (b) Immunofluorescence staining of H3K27me3 after TGF-β or TGF-β with GSK-126 exposure (a.u. Represents arbitrary units) (Scale bar: 10 μm). White arrow denotes H3K27me3 protein. (c and d) Transcript levels of GOT1 (c), CCNA2, CCNB1 (d) and cell cycle distribution post TGF-β exposure with or without GSK-126 (e). (f) Protein level of PCNA after TGF-β or TGF-β plus GSK-126 exposure. (g and h) Protein expression of EZH2 (g) and transcript levels of GOT1, CCNA2 and CCNB1 after TGF-β or TGF-β with EZH2-siRNA exposure (h). β-Actin acted as loading control for immunoblots. Wherever applicable, TGF-β was at a dose of 5 ng/ml for 3 days. Data were statistically analysed using results obtained from minimum three or more replicates. The p-value significance was estimated by two way- ANOVA and unpaired two-tailed t-tests. Uncropped images of blots are shown in Figure S10-4 (a, f, g).

Article Snippet: Other reagents used were the Fetal Bovine Serum (FBS; Invitrogen, 26,140–079), Enhanced Chemiluminescence kit (ECL; Thermo Scientific, 32,106), GSK-126 (Merck, 500,580), Signal Silence EZH2siRNAi (CST, 6509), SMAD inhibitor- SIS3 (BioVision, 2227), magnetic beads (BioRad; 161–4013), Lipofectamine (Invitrogen, RNAiMax), and Dulbecco's modified minimal essential culture medium (DMEM; Gibco, 12,800–017).

Techniques: Immunofluorescence, Staining, Expressing, Western Blot, Two Tailed Test

Journal: Heliyon

Article Title: Transforming growth factor- β mediated regulation of epigenome is required for epithelial to mesenchymal transition associated features in liver cancer cells

doi: 10.1016/j.heliyon.2023.e14665

Figure Lengend Snippet: EZH2 inhibition reverses EMT by suppressing SMADs. (a–e) N-Cad and Vimentin protein (a) and RNA levels (b), immunoblot of E-Cad (c) and SMAD2/3 (d) and immunofluorescence staining of SMAD2 (e) after TGF-β with or without GSK-126 treatment (Scale bar: 10 μm) (a.u. Represents arbitrary units). White arrow denotes SMAD2 protein. (f and g) Protein expression of SMAD2/3 (f) and Vimentin (g) after TGF-β or TGF-β with EZH2-siRNA. (h) Immunofluorescence staining of Vimentin after TGF-β or TGF-β with GSK-126 or TGF-β with EZH2-siRNA treatment (Scale bar: 10 μm). Vimentin protein is marked by white arrow. Relative intensity for immunoblots was measured and quantified using ImageJ. β-Actin acted as loading control for immunoblots. Wherever applicable, TGF-β was at a dose of 5 ng/ml for 3 days. Data were statistically analysed using results obtained from minimum three or more replicates. The p-value significance was estimated by two way- ANOVA and unpaired two-tailed t-tests. Uncropped images of blots are shown in Figure S13-7 (a, c) & Figure S14-7 (d, f, g).

Article Snippet: Other reagents used were the Fetal Bovine Serum (FBS; Invitrogen, 26,140–079), Enhanced Chemiluminescence kit (ECL; Thermo Scientific, 32,106), GSK-126 (Merck, 500,580), Signal Silence EZH2siRNAi (CST, 6509), SMAD inhibitor- SIS3 (BioVision, 2227), magnetic beads (BioRad; 161–4013), Lipofectamine (Invitrogen, RNAiMax), and Dulbecco's modified minimal essential culture medium (DMEM; Gibco, 12,800–017).

Techniques: Inhibition, Western Blot, Immunofluorescence, Staining, Expressing, Two Tailed Test