Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Targeting CDK2 Confers Vulnerability to Lenvatinib Via Driving Senescence in Anaplastic Thyroid Cancer.

doi: 10.1002/advs.202413514

Figure Lengend Snippet: Figure 2. Combined inhibition effect of lenvatinib with CDK2 suppression in ATC. a) Western blot analyses of CDK1, CDK2, CDK5, CDK9 in KHM-5M, C643 and PDC13 transfected with siRNAs against CDK1, CDK2, CDK5, CDK9 and negative control. b-d) KHM-5M (b), C643 (c) and PDC13 (d) were transfected with siRNAs against CDK1, CDK2, CDK5, CDK9 and negative control, followed by cell viability testing after treatment with lenvatinib (10, 20, and 40 μM). Relative cell viability is calculated as the percentage relative to the cell viability of cells treated with lenvatinib = 0 μM in each group.

Article Snippet: The following antibodies were used for western blot: Tubulin (1:1000, #2146, Cell Signaling Technology), CDK1 (1:1000, #77055, Cell Signaling Technology), CDK2 (1:1000, #18048, Cell Signaling Technology), CDK5 (1:1000, #2506, Cell Signaling Technology), CDK9 (1:1000, #2316, Cell Signaling Technology), p21 (1:1000, #2947, Cell Signaling Technology), p16 (1:1000, #80772, Cell Signaling Technology), TP53 (1:20000, 60283-2-lg, Proteintech), Lamin B1 (1:5000, 66095-1-lg, Proteintech), p-CDK2 (Thr160) (1:1000, #2561, Cell Signaling Technology), Rb (1:2000, #9309, Cell Signaling Technology), p-Rb (Ser 807/811) (1:1000, #8516, Cell Signaling Technology), cleaved caspase-3 (1:1000, #25128-1-AP, Proteintech), ubiquitin (1:1000, #10201- 2-AP, Proteintech), Flag (1:1000, #F3165, Sigma-Aldrich), Myc (1:5000, #60003-2-Ig, Proteintech), HA (1:5000, #51064-2-AP, Proteintech), RACK1 (1:1000, #5432, Cell Signaling Technology).

Techniques: Inhibition, Western Blot, Transfection, Negative Control

Journal: Journal of Biological Chemistry

Article Title: Large T Antigen Promotes JC Virus Replication in G2-arrested Cells by Inducing ATM- and ATR-mediated G2 Checkpoint Signaling

doi: 10.1074/jbc.m109.064311

Figure Lengend Snippet: FIGURE 7. Suppression of JCV genome replication by abrogation of G2 arrest. Effect of inhibition of G2 check- point signaling on JCV DNA replication as determined by the DpnI replication assay. IMR-32 cells transfected with pBS-JCori and an expression vector for TAg were incubated for 24 h before addition of dimethyl sulfoxide (DMSO) (control), 50 nM ucn-01, or 2.5 mM caffeine (caf). The siRNA against non-targeting control (Ct) or Wee1 or/and Cdc2 were transfected 24 h before transfection of the plasmids. The collected cells were subjected to immunoblot anal- ysis with antibodies to FLAG-TAg, Wee1, and Cdc2 (A), flow cytometric analysis of cell cycle profile for the TAg cell subset (B), and DpnI replication assay (C and D). Replicated DNA extracted from the cells was detected by Southern blotanalysiswithaDNAprobespecificforpBS-JCori(C).TheintensityofDNAbandswasquantifiedandisindicated inthebargraphrelativetothevalueofdimethylsulfoxide(forucn-01andcaffeine)orthatofsiCt(forsiWee1,siCdc2, and siWeesiCdc2) (D); data are mean S.D. of triplicates from a representative experiment.

Article Snippet: Antibodies to Chk1, Ser317-phosphorylated Chk1, Chk2, Thr68-phosphorylated Chk2, Tyr15-phosphorylated Cdc2, and Ser15-phosphorylated p53 were obtained from Cell Signaling Technology (Beverly,MA).

Techniques: Inhibition, Transfection, Expressing, Plasmid Preparation, Incubation, Control, Western Blot, FLAG-tag