mouse monoclonal antibodies against cdk6  (Cell Signaling Technology Inc)

Journal: Frontiers in Cell and Developmental Biology

Article Title: Recreating the Bone Marrow Microenvironment to Model Leukemic Stem Cell Quiescence

doi: 10.3389/fcell.2021.662868

Figure Lengend Snippet: Hypoxia and co-culture with bone marrow mesenchymal stromal cells (BMSC) + TGFβ-1 increases the expression of markers of leukemia cell quiescence. KG1a cells were cultured alone or with alginate-encapsulated BMSC and TGFβ-1 (10 ng/ml) over a 12-day period in normoxia or hypoxia (1% pO 2 ). (A) Expression of CD150 determined in the live fraction by flow cytometry. MFI, mean fluorescent intensity. * p < 0.05 (Student’s t -test). (B–E) Results of quantitative RT-PCRs on RNA isolated on days 8 and 12. * p < 0.05 (one-way ANOVA). (F) Gene expression analysis of p21 expression conducted on the GSE17054 transcriptomic dataset. The graph represents the changes in gene expression between hematopoietic stem cells and leukemic stem cells. * p < 0.05 (Welch’s t -test). (G) Western blot images for the expression of p21, CDK4, and CDK6 conducted on samples isolated at day 12 of culture. SN, single culture at normoxia; CH, co-culture under hypoxia.

Article Snippet: After blocking, the blots were incubated with rabbit monoclonal antibodies against CDK4 [Cell Signaling Technology (CST)] and p21 (Santa Cruz) and with mouse monoclonal antibodies against CDK6 (CST) and actin (Sigma-Aldrich).

Techniques: Co-Culture Assay, Expressing, Cell Culture, Flow Cytometry, Isolation, Gene Expression, Western Blot

Journal: Biochemical pharmacology

Article Title: Mechanisms of tolvaptan-induced toxicity in HepG2 cells ☆

doi: 10.1016/j.bcp.2015.03.015

Figure Lengend Snippet: HepG2 cells were treated with tolvaptan (0–100 μM) for 24 or 48 h. (A) Cell cycle progression was determined as described in Materials and Methods. The results shown are the mean and standard deviation of three independent experiments. * Significantly (p < 0.05) different from 0 μM tolvaptan. (B) Percentage of p-H3 positive cells. The results shown are the mean and standard deviation of three independent experiments. * Significantly (p < 0.05) different from 0 μM tolvaptan. (C) Representative Western blot images of cyclin D1, cyclin D3, cyclin B1, CDK1, CDK2, CDK4, and CDK6 in the HepG2 cells treated with tolvaptan. β-Actin was used as a loading control.

Article Snippet: Mouse monoclonal antibodies against cyclin D1, cyclin D3, cyclin-dependent kinase (CDK) 4 (CDK4), and CDK6, rabbit monoclonal antibodies against CDK2, phosphorylated checkpoint kinase 1 at Ser345 (p-Chk1), Bcl-2, Bad, Bim, Bax, phosphorylated-ERK1/2 at Thr202/Tyr204 (p-ERK1/2), ERK1/2, phosphorylated-JNK at Thr183/Tyr185 (p-JNK), JNK, phosphorylated-p38 at Thr180/Tyr182 (p-p38), and p38, and rabbit polyclonal antibodies against phosphorylated checkpoint kinase 2 at Thr68 (p-Chk2), cytochrome C, and Bid were purchased from Cell Signaling Technology (Danvers, MA).

Techniques: Standard Deviation, Western Blot, Control

Journal: Biochemical pharmacology

Article Title: Mechanisms of tolvaptan-induced toxicity in HepG2 cells ☆

doi: 10.1016/j.bcp.2015.03.015

Figure Lengend Snippet: HepG2 cells were treated with tolvaptan (0, 40, or 80 μM) in the presence or absence of the proteasome inhibitor MG-132 (100 nM) for 24 h. (A) Representative Western blot images of cyclin D1, cyclin D3, CDK1, CDK2, CDK4, and CDK6. β-Actin was used as a loading control. (B) Cell cycle distribution. The results shown are the mean and standard deviation of three independent experiments. * Significantly (p < 0.05) different from cells without MG-132 treatment.

Article Snippet: Mouse monoclonal antibodies against cyclin D1, cyclin D3, cyclin-dependent kinase (CDK) 4 (CDK4), and CDK6, rabbit monoclonal antibodies against CDK2, phosphorylated checkpoint kinase 1 at Ser345 (p-Chk1), Bcl-2, Bad, Bim, Bax, phosphorylated-ERK1/2 at Thr202/Tyr204 (p-ERK1/2), ERK1/2, phosphorylated-JNK at Thr183/Tyr185 (p-JNK), JNK, phosphorylated-p38 at Thr180/Tyr182 (p-p38), and p38, and rabbit polyclonal antibodies against phosphorylated checkpoint kinase 2 at Thr68 (p-Chk2), cytochrome C, and Bid were purchased from Cell Signaling Technology (Danvers, MA).

Techniques: Western Blot, Control, Standard Deviation