Journal:

Article Title: Mitotic clonal expansion: A synchronous process required for adipogenesis

doi: 10.1073/pnas.0137044100

Figure Lengend Snippet: Effect of PD98059, a MEK inhibitor, on MAPK kinase activity, cell cycle markers, MCE, and adipogenesis. (A) Two-day postconfluent 3T3-L1 preadipocytes were induced to differentiate by using the standard differentiation protocol. At various time points, cell lysates were prepared and subjected to SDS/PAGE and immunoblotting with antibodies to MAPK and phospho-MAPK. (B) Two-day postconfluent 3T3-L1 preadipocytes were exposed to 20 or 50 μM PD98059 for 30 min before induction and again at the time of induction. Cell lysates were prepared 1 h later for MAPK and phospho-MAPK and 20 h later for cyclin A and cdk2 immunoblot analyses. (C) Two-day postconfluent 3T3-L1 preadipocytes were exposed to 20 or 50 μM PD98059 for 30 min before induction and again at the time of induction. On days 2 and 4, cell number was determined. MDI refers to the differentiation inducers. (D) Postconfluent 3T3-L1 preadipocytes were treated as in B. Cell lysates were prepared on day 3, and 50 μg of lysate protein were subjected to SDS/PAGE and immunoblotted with antibodies against C/EBPα, PPARγ, and 422/aP2. (E) Cells were treated as in B, above, except that cell lysates were prepared on day 6. (F) Cells were treated as B, except that on day 8, cells were fixed and cytoplasmic triacylglycerol was stained with Oil Red-O. PD98059 concentrations are given (20 and 50 μM).

Article Snippet: Whereas most cell types express cdk2 throughout the cell cycle including G 1 ( 32 ), 3T3-L1 preadipocytes do not express cdk2 in the G 1 phase of MCE (Fig. C ).

Techniques: Activity Assay, SDS Page, Western Blot, Staining

Journal:

Article Title: Mitotic clonal expansion: A synchronous process required for adipogenesis

doi: 10.1073/pnas.0137044100

Figure Lengend Snippet: Changes in the expression and translocation of cell-cycle markers and GSK-3β during MCE. (A) Day 0 postconfluent 3T3-L1 preadipocytes were induced to differentiate into adipocytes. At the times indicated, whole-cell lysates (50 μg of protein) were subjected to SDS/PAGE and immunoblotted. The decreased mobility of Rb after 16 h was shown to be caused by hyperphosphorylation by incubating nuclear extracts with alkaline phosphatase, which increased the mobility of Rb (not shown). (B) 3T3-L1 preadipocytes on glass coverslips were cultured to postconfluence in DMEM containing 10% calf serum and induced to differentiate. At 8 and 20 h, cells were fixed and exposed to cyclin D1 antibody followed by FITC-labeled secondary antibody. (C) At the indicated times after induction of differentiation, cell lysates (50 μg of protein) were subjected to SDS/PAGE immunoblotted with antibodies to cdk2 and cdk4. (D) Nuclear extracts were prepared at the indicated times after induction. Cdk2 was immunoprecipitated and incubated with purified histone H1 and [γ-32P]ATP. After SDS/PAGE, [32P]histone H1 was detected by autoradiography. Total histone H1 was detected by Coomassie blue staining. (E) At the indicated times after induction of differentiation, preadipocytes were lysed and resolved into nuclear and cytoplasmic fractions. The whole-cell lysate and nuclear fractions (identical cell equivalents) were subjected to SDS/PAGE and then immunoblotted with antibody to GSK-3β.

Article Snippet: Whereas most cell types express cdk2 throughout the cell cycle including G 1 ( 32 ), 3T3-L1 preadipocytes do not express cdk2 in the G 1 phase of MCE (Fig. C ).

Techniques: Expressing, Translocation Assay, SDS Page, Cell Culture, Labeling, Immunoprecipitation, Incubation, Purification, Autoradiography, Staining

Journal:

Article Title: Mitotic clonal expansion: A synchronous process required for adipogenesis

doi: 10.1073/pnas.0137044100

Figure Lengend Snippet: Effect of U0126 (a MEK inhibitor) on MCE, expression of cell cycle and adipocyte markers, and adipogenesis. (A) Two-day postconfluent 3T3-L1 preadipocytes were incubated with 20 or 40 μM U0126 for 30 min and then induced to differentiate with the same amount of U0126; U0126 was added again 24 h later. Cell lysates were prepared 1 h later for MAPK and phospho-MAPK analysis and at 20 h for cyclin A and cdk2 analysis. Cell lysates (50 μg of protein) were subjected to SDS/PAGE and immunoblotting as described in Fig. ​Fig.3.3. (B) Two-day postconfluent 3T3-L1 preadipocytes were treated as A, and cell number was determined on day 4. (C) Two-day postconfluent 3T3-L1 preadipocytes were treated as in A; cell lysates were prepared on day 6 and subjected to analysis as in A with antibodies to C/EBPα, PPARγ, and 422/aP2. (D) Two-day postconfluent 3T3-L1 preadipocytes were treated as in A; on day 8, cells were fixed, and cytoplasmic triacylglycerol was stained with Oil Red-O. U0126 concentrations are given (20 and 40 μM).

Article Snippet: Whereas most cell types express cdk2 throughout the cell cycle including G 1 ( 32 ), 3T3-L1 preadipocytes do not express cdk2 in the G 1 phase of MCE (Fig. C ).

Techniques: Expressing, Incubation, SDS Page, Western Blot, Staining

Journal: Cancer medicine

Article Title: The expression and role of SUZ12 in lung adenocarcinoma.

doi: 10.1002/cam4.70190

Figure Lengend Snippet: FIGURE 7 The effect of SUZ12 on CDKs and cyclins expression was tested by qRT-PCR and western blotting. sh-SUZ12 decreased CDK3 mRNA (A), CDK2/3 (C), and cyclin D1 (F) protein expression, while increased cyclin E1 protein expression (F), without significantly effected the cyclins mRNA (E). oe-SUZ12 increased CDK3/6 mRNA (B) and CDK3 protein expression (D). *p < 0.05, **p < 0.001.

Article Snippet: The list of primary antibodies: SUZ12 (1 μg/mL, Abcam Cambridge, cat no: ab12073), CDK2 (1:1000; Proteintech, USA, cat. no. 10122- 1- AP), CDK3 (1:2000; Proteintech, USA, cat. no. 55103- 1- AP), CDK6 (1:2000; Proteintech, USA, cat. no. 14052- 1- AP), cyclin D1 (1:5000; Proteintech, USA, cat. no. 26939- 1- AP), cyclin E1 (1:1000; Proteintech, USA, cat. no. 11554- 1- AP), p18 (1:1000, BOSTER China, cat. no. M03299- 1), p19 (1:1000, BOSTER China, cat. no. MA1075), p53 (1:5000; Proteintech, USA, cat. no. 60283- 2- Ig), p- p53 (1:2000; Proteintech, USA, cat. no. 28961- 1- AP), p57 (1:1000, BOSTER China, cat. no. BM4129), Rb (1:1000, BOSTER China, cat. no. BM4500), pRb (1:1000, BOSTER China, cat. no. BM4338), Bcl- 2 (1:1000; Proteintech, USA, cat. no. 26593- 1- AP), Bax (1:2000; Proteintech, USA, cat. no. 50599- 2- lg), Ecadherin (1:5000; Proteintech, USA, cat. no. 20874- 1- AP), N- cadherin (1:3000; Proteintech, USA, cat. no. 22018- 1- AP), vimentin (1:4000; Proteintech, USA, cat. no. 10366- 1- AP), MMP1 (1:1000, BOSTER China, cat. no. A00733- 1), MMP2 (1:500, BOSTER China, cat. no. BM4075), MMP9 (1:1000, BOSTER China, cat. no. PB0709), MMP14 (1:1000, BOSTER China, cat. no. BM4119), TIMP1 (1:1000, Bioss China, cat. no. bs0415R), TIMP2 (1:1000, Bioss China, cat. no. bs- 10395R), TIMP3 (1:1000; Proteintech, USA, cat. no. 10858- 1- AP), ITGB1 (1:1000, BOSTER China, cat. no. BM4308), ITGB3 (1:1000, BOSTER China, cat. no. BA1670), ITGB5 (1:1000, BOSTER China, cat. no. A04201- 1), nm23 (1:1000, BOSTER China, cat. no. BA3787), PD- L1 (1:3000; Proteintech, USA, cat. no. 66248- 1- Ig), and βactin (1:5000; Proteintech, USA, cat. no. 66009- 1- Ig).

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: NT157 inhibits cell proliferation and sensitizes glioma cells to TRAIL-induced apoptosis by up-regulating DR5 expression.

doi: 10.1016/j.biopha.2022.113502

Figure Lengend Snippet: Fig. 8. Proposed signal pathways. NT157 caused ROS generation and DNA damage, which activated p21 and p27, and subsequently lunched S-phase and M-phase cell cycle arrest through regulating cyclin A1, CDK2, cyclin B1 and CDK1. NT157 also dysfunctioned MAPKs, PI3K/AKT and EGFR-STAT3 pathways, and enhanced TRAIL-induced glioma cells apoptosis by up-regulating DR5 expression.

Article Snippet: CyclinA (sc-271682), anti-p21(sc-6246), CDK1 (sc-53219) antibodies were purchased from Santa Cruz Biotechnology (Shanghai, China). β-actin antibody and IgG were bought from Beijing zhongshan Jinqiao Biotechnology (Beijing, China). p27 (#3686), CDK2 (#18048), Cyclin B1 (#12231), PARP (#9542), Caspase-3 (#9662), PCNA (#13110), Ku70 (#4588), Ku80 (#2753), γ-H2AX (#80312), p-ERK (#4370), ERK (#4695), p-p38 (#4511), p38 (#8690), p-JNK (#9255), JNK (#9252), Akt (#4685), p-Akt (#4060), EGFR (#8083), p-Stat3 (#9145), Stat3 (#9139), DR5(#69400), DR4(#42533) were all purchased from Cell Signaling Technology (CST, USA).

Techniques: Expressing

Journal: Pharmaceuticals

Article Title: Synthesis and Evaluation of Some New 4 H -Pyran Derivatives as Antioxidant, Antibacterial and Anti-HCT-116 Cells of CRC, with Molecular Docking, Antiproliferative, Apoptotic and ADME Investigations

doi: 10.3390/ph15070891

Figure Lengend Snippet: Examples of pyran-containing compounds with antibacterial , antioxidant and anticancer activities via inhibition of CDK2 [ , , ].

Article Snippet: CDK-2 inhibition activity of the studied compounds 4d and 4k was evaluated using a commercial CDK2 ELISA Kit (cat. no.: 79599; BPS Biosciences, San Diego, CA, USA) following the manufacturer’s instructions.

Techniques: Inhibition

Journal: Pharmaceuticals

Article Title: Synthesis and Evaluation of Some New 4 H -Pyran Derivatives as Antioxidant, Antibacterial and Anti-HCT-116 Cells of CRC, with Molecular Docking, Antiproliferative, Apoptotic and ADME Investigations

doi: 10.3390/ph15070891

Figure Lengend Snippet: Redocked (yellow) and co-crystallized (baby blue) ligand ( DTQ ) in the ATP binding pocket of CDK2 after self-docking calculations.

Article Snippet: CDK-2 inhibition activity of the studied compounds 4d and 4k was evaluated using a commercial CDK2 ELISA Kit (cat. no.: 79599; BPS Biosciences, San Diego, CA, USA) following the manufacturer’s instructions.

Techniques: Binding Assay

Journal: Pharmaceuticals

Article Title: Synthesis and Evaluation of Some New 4 H -Pyran Derivatives as Antioxidant, Antibacterial and Anti-HCT-116 Cells of CRC, with Molecular Docking, Antiproliferative, Apoptotic and ADME Investigations

doi: 10.3390/ph15070891

Figure Lengend Snippet: Binding free energy of 4d , 4k , 4f , BMS-265246 and DTQ at the active site of CDK2.

Article Snippet: CDK-2 inhibition activity of the studied compounds 4d and 4k was evaluated using a commercial CDK2 ELISA Kit (cat. no.: 79599; BPS Biosciences, San Diego, CA, USA) following the manufacturer’s instructions.

Techniques: Binding Assay

Journal: Pharmaceuticals

Article Title: Synthesis and Evaluation of Some New 4 H -Pyran Derivatives as Antioxidant, Antibacterial and Anti-HCT-116 Cells of CRC, with Molecular Docking, Antiproliferative, Apoptotic and ADME Investigations

doi: 10.3390/ph15070891

Figure Lengend Snippet: Three-dimensional binding interactions of DTQ within the ATP binding pocket of CDK2. Hydrogen bonds (yellow dotted lines), hydrogen atoms (white), nitrogen atoms (blue), and oxygen atoms (red).

Article Snippet: CDK-2 inhibition activity of the studied compounds 4d and 4k was evaluated using a commercial CDK2 ELISA Kit (cat. no.: 79599; BPS Biosciences, San Diego, CA, USA) following the manufacturer’s instructions.

Techniques: Binding Assay

Journal: Pharmaceuticals

Article Title: Synthesis and Evaluation of Some New 4 H -Pyran Derivatives as Antioxidant, Antibacterial and Anti-HCT-116 Cells of CRC, with Molecular Docking, Antiproliferative, Apoptotic and ADME Investigations

doi: 10.3390/ph15070891

Figure Lengend Snippet: Two-dimensional binding interactions of DTQ within the ATP binding pocket of CDK2.

Article Snippet: CDK-2 inhibition activity of the studied compounds 4d and 4k was evaluated using a commercial CDK2 ELISA Kit (cat. no.: 79599; BPS Biosciences, San Diego, CA, USA) following the manufacturer’s instructions.

Techniques: Binding Assay

Journal: Pharmaceuticals

Article Title: Synthesis and Evaluation of Some New 4 H -Pyran Derivatives as Antioxidant, Antibacterial and Anti-HCT-116 Cells of CRC, with Molecular Docking, Antiproliferative, Apoptotic and ADME Investigations

doi: 10.3390/ph15070891

Figure Lengend Snippet: Three-dimensional binding interactions of BMS-265246 within the ATP binding pocket of CDK2. Hydrogen bond (purple dotted line), hydrogen atoms (white), nitrogen atoms (blue), fluorine atoms (light green), and oxygen atoms (red).

Article Snippet: CDK-2 inhibition activity of the studied compounds 4d and 4k was evaluated using a commercial CDK2 ELISA Kit (cat. no.: 79599; BPS Biosciences, San Diego, CA, USA) following the manufacturer’s instructions.

Techniques: Binding Assay

Journal: Pharmaceuticals

Article Title: Synthesis and Evaluation of Some New 4 H -Pyran Derivatives as Antioxidant, Antibacterial and Anti-HCT-116 Cells of CRC, with Molecular Docking, Antiproliferative, Apoptotic and ADME Investigations

doi: 10.3390/ph15070891

Figure Lengend Snippet: Two-dimensional interactions of BMS-265246 within the ATP binding pocket of CDK2 kinase.

Article Snippet: CDK-2 inhibition activity of the studied compounds 4d and 4k was evaluated using a commercial CDK2 ELISA Kit (cat. no.: 79599; BPS Biosciences, San Diego, CA, USA) following the manufacturer’s instructions.

Techniques: Binding Assay

Journal: Pharmaceuticals

Article Title: Synthesis and Evaluation of Some New 4 H -Pyran Derivatives as Antioxidant, Antibacterial and Anti-HCT-116 Cells of CRC, with Molecular Docking, Antiproliferative, Apoptotic and ADME Investigations

doi: 10.3390/ph15070891

Figure Lengend Snippet: Three-dimensional model of binding interactions of compound 4d after docking calculations in the ATP binding pocket of CDK2. Hydrogen bond (black lines), hydrogen atoms (white), nitrogen atoms (blue), and oxygen atoms (red).

Article Snippet: CDK-2 inhibition activity of the studied compounds 4d and 4k was evaluated using a commercial CDK2 ELISA Kit (cat. no.: 79599; BPS Biosciences, San Diego, CA, USA) following the manufacturer’s instructions.

Techniques: Binding Assay

Journal: Pharmaceuticals

Article Title: Synthesis and Evaluation of Some New 4 H -Pyran Derivatives as Antioxidant, Antibacterial and Anti-HCT-116 Cells of CRC, with Molecular Docking, Antiproliferative, Apoptotic and ADME Investigations

doi: 10.3390/ph15070891

Figure Lengend Snippet: Two-dimensional model of binding interactions of compound 4d after docking calculations in the ATP binding pocket of CDK2.

Article Snippet: CDK-2 inhibition activity of the studied compounds 4d and 4k was evaluated using a commercial CDK2 ELISA Kit (cat. no.: 79599; BPS Biosciences, San Diego, CA, USA) following the manufacturer’s instructions.

Techniques: Binding Assay

Journal: Pharmaceuticals

Article Title: Synthesis and Evaluation of Some New 4 H -Pyran Derivatives as Antioxidant, Antibacterial and Anti-HCT-116 Cells of CRC, with Molecular Docking, Antiproliferative, Apoptotic and ADME Investigations

doi: 10.3390/ph15070891

Figure Lengend Snippet: Three-dimensional model of binding interactions of compound 4K after docking calculations in the ATP binding pocket of CDK2. Hydrogen bond (black lines), hydrogen atoms (white), nitrogen atoms (blue), and oxygen atoms (red).

Article Snippet: CDK-2 inhibition activity of the studied compounds 4d and 4k was evaluated using a commercial CDK2 ELISA Kit (cat. no.: 79599; BPS Biosciences, San Diego, CA, USA) following the manufacturer’s instructions.

Techniques: Binding Assay

Journal: Pharmaceuticals

Article Title: Synthesis and Evaluation of Some New 4 H -Pyran Derivatives as Antioxidant, Antibacterial and Anti-HCT-116 Cells of CRC, with Molecular Docking, Antiproliferative, Apoptotic and ADME Investigations

doi: 10.3390/ph15070891

Figure Lengend Snippet: Two-dimensional model of binding interactions of compound 4K after docking calculations in the ATP binding pocket of CDK2.

Article Snippet: CDK-2 inhibition activity of the studied compounds 4d and 4k was evaluated using a commercial CDK2 ELISA Kit (cat. no.: 79599; BPS Biosciences, San Diego, CA, USA) following the manufacturer’s instructions.

Techniques: Binding Assay

Journal: Pharmaceuticals

Article Title: Synthesis and Evaluation of Some New 4 H -Pyran Derivatives as Antioxidant, Antibacterial and Anti-HCT-116 Cells of CRC, with Molecular Docking, Antiproliferative, Apoptotic and ADME Investigations

doi: 10.3390/ph15070891

Figure Lengend Snippet: Three-dimensional model of binding interactions of compound 4f after docking calculations in the ATP binding pocket of CDK2. Hydrogen atoms (white), nitrogen atoms (blue), chlorine atom (green), and oxygen atoms (red).

Article Snippet: CDK-2 inhibition activity of the studied compounds 4d and 4k was evaluated using a commercial CDK2 ELISA Kit (cat. no.: 79599; BPS Biosciences, San Diego, CA, USA) following the manufacturer’s instructions.

Techniques: Binding Assay

Journal: Pharmaceuticals

Article Title: Synthesis and Evaluation of Some New 4 H -Pyran Derivatives as Antioxidant, Antibacterial and Anti-HCT-116 Cells of CRC, with Molecular Docking, Antiproliferative, Apoptotic and ADME Investigations

doi: 10.3390/ph15070891

Figure Lengend Snippet: Two-dimensional model of binding interactions of compound 4f after docking calculations in the ATP binding pocket of CDK2.

Article Snippet: CDK-2 inhibition activity of the studied compounds 4d and 4k was evaluated using a commercial CDK2 ELISA Kit (cat. no.: 79599; BPS Biosciences, San Diego, CA, USA) following the manufacturer’s instructions.

Techniques: Binding Assay

Journal: Pharmaceuticals

Article Title: Synthesis and Evaluation of Some New 4 H -Pyran Derivatives as Antioxidant, Antibacterial and Anti-HCT-116 Cells of CRC, with Molecular Docking, Antiproliferative, Apoptotic and ADME Investigations

doi: 10.3390/ph15070891

Figure Lengend Snippet: In vitro evaluation of the CDK2 inhibitory efficiency of pyrans 4d and 4K as compared to the reference inhibitor BMS-265246 over a concentration range of 0.01−10 µM. Results were obtained from three independent experiments.

Article Snippet: CDK-2 inhibition activity of the studied compounds 4d and 4k was evaluated using a commercial CDK2 ELISA Kit (cat. no.: 79599; BPS Biosciences, San Diego, CA, USA) following the manufacturer’s instructions.

Techniques: In Vitro, Concentration Assay

Journal: Pharmaceuticals

Article Title: Synthesis and Evaluation of Some New 4 H -Pyran Derivatives as Antioxidant, Antibacterial and Anti-HCT-116 Cells of CRC, with Molecular Docking, Antiproliferative, Apoptotic and ADME Investigations

doi: 10.3390/ph15070891

Figure Lengend Snippet: IC 50 values of 4d , 4K and BMS-265246 against CDK2.

Article Snippet: CDK-2 inhibition activity of the studied compounds 4d and 4k was evaluated using a commercial CDK2 ELISA Kit (cat. no.: 79599; BPS Biosciences, San Diego, CA, USA) following the manufacturer’s instructions.

Techniques:

Journal: Pharmaceuticals

Article Title: Synthesis and Evaluation of Some New 4 H -Pyran Derivatives as Antioxidant, Antibacterial and Anti-HCT-116 Cells of CRC, with Molecular Docking, Antiproliferative, Apoptotic and ADME Investigations

doi: 10.3390/ph15070891

Figure Lengend Snippet: In vitro quantitative determination of the concentrations of CDK2 (ng/mL) in HCT116 cells treated with pyrans 4d and 4K compared with the positive control BMS-265246 and the negative control samples.

Article Snippet: CDK-2 inhibition activity of the studied compounds 4d and 4k was evaluated using a commercial CDK2 ELISA Kit (cat. no.: 79599; BPS Biosciences, San Diego, CA, USA) following the manufacturer’s instructions.

Techniques: In Vitro, Positive Control, Negative Control

Journal: Pharmaceuticals

Article Title: Synthesis and Evaluation of Some New 4 H -Pyran Derivatives as Antioxidant, Antibacterial and Anti-HCT-116 Cells of CRC, with Molecular Docking, Antiproliferative, Apoptotic and ADME Investigations

doi: 10.3390/ph15070891

Figure Lengend Snippet: The expression profiles of the CDK2 gene in the lysate of HCT-116 cancer cells treated with compounds 4d and 4 k .

Article Snippet: CDK-2 inhibition activity of the studied compounds 4d and 4k was evaluated using a commercial CDK2 ELISA Kit (cat. no.: 79599; BPS Biosciences, San Diego, CA, USA) following the manufacturer’s instructions.

Techniques: Expressing