anti cyclin e2  (Proteintech)

Journal: Ecotoxicology and environmental safety

Article Title: The influence of phenolic environmental estrogen on the transcriptome of uterine leiomyoma cells: A whole transcriptome profiling-based analysis.

doi: 10.1016/j.ecoenv.2021.111945

Figure Lengend Snippet: Fig. 4. The effects of 10.0 μmol/L BPA, 32.0 μmol/L NP on genes and proteins expression in uterine leiomyoma cells. (a) Hierarchically clustered heatmap of differentially expressed genes associated with cell cycle after BPA and NP exposure. (b) E2F1, CCND1, CCNE2, MCM2, MCM3, MCM4, MCM5, MCM6 genes expression by q-PCR presented as relative changes. There were increased E2F1, CCND1, CCNE2, MCM2, MCM3, MCM4, MCM5, MCM6 genes expression in uterine leiomyoma cells treated with10.0 μmol/L BPA, 32.0 μmol/L NP for 48 h compared to control group (*P < 0.05). (c) Representative western blots and (d) band intensity bar graphs of proteins. There were significantly (*P < 0.05) higher expression levels of E2F1, CCND1, CCNE2, MCM2, MCM3, MCM4, MCM5, MCM6 in the cells treated with BPA and NP for 48 h compared to controls. The western band images shown are representative of three independent experiments and the data were expressed as mean ± SE done in three independent experiments.

Article Snippet: The following primary antibodies were used for the western blotting: E2F1 Monoclonal Antibody (66515-1-Ig; Proteintech; diluted 1:1000); Cyclin D1 Monoclonal Antibody (60186-1-Ig; Proteintech; diluted 1:5000); Cyclin E2 Polyclonal Antibody (11935-1-AP; Proteintech; diluted 1:500); MCM2 Polyclonal Antibody (10513-1-AP; Proteintech; diluted 1:1500); MCM3 Polyclonal Antibody (15597-1-AP; Proteintech; diluted 1:1000); MCM4 Polyclonal Antibody (13043-1-AP; Proteintech; diluted 1:600); MCM5 Polyclonal Antibody (11703-1-AP; Proteintech; diluted 1:1000); MCM6 Polyclonal Antibody (13347-2-AP; Proteintech; diluted 1:8000); phospho-Rb monoclonal (ab184796; abcam; diluted 1:1000); phospho-PI3K polyclonal (ab182651; abcam; diluted 1:500); phospho- AKT polyclonal (ab38449; abcam; diluted 1:500).

Techniques: Expressing, Control, Western Blot

Journal: Molecular and cellular endocrinology

Article Title: DUAL INHIBITION OF ERK1/2 AND AKT PATHWAYS IS REQUIRED TO SUPPRESS THE GROWTH AND SURVIVAL OF ENDOMETRIOTIC CELLS AND LESIONS

doi: 10.1016/j.mce.2018.12.011

Figure Lengend Snippet: Details of the antibody used.

Article Snippet: Anti-human rabbit polyclonal Cyclin E2 , Cell Signaling , 4132 , 1:1000.

Techniques: Concentration Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Motor neuron and pancreas homeobox 1/HLXB9 promotes sustained proliferation in bladder cancer by upregulating CCNE1/2

doi: 10.1186/s13046-018-0829-9

Figure Lengend Snippet: MNX1 directly upregulates CCNE1 and CCNE2 promoter activity in bladder cancer cells. a Real-time PCR of cell cycle–related gene mRNA expression. Gene expression levels were normalized to GAPDH . Pseudo-colors represent the intensity scale of MNX1 versus vector or MNX1 shRNA#1/2 versus Scramble, generated by log2 transformation. b Western blotting of CCNE1, CCNE2, phosphorylated Rb (p-Rb), and total Rb protein expression; α-tubulin was used as the loading control. c The scatter diagram of CCNE1 and CCNE2 from cBioportal program. d and e Left: Luciferase activity assays of T24 and 5637 cells showed transactivation and repression of the CCNE1 ( d ) and CCNE2 ( e ) promoters by MNX1 overexpression and MNX1 knockdown, respectively. Right: Schematic illustration of ChIP PCR fragments for the indicated nucleotide regions of the CCNE1 ( d ) and CCNE2 ( e ) promoters (top). The ChIP enrichment assay confirmed that MNX1 binds to the P4 and P5 promoter of CCNE1 ( d ) and the P6 and P7 promoters of CCNE2 ( e ); immunoglobulin G (IgG) was used as the negative control. The results from three independent experiments were evaluated. * p < 0.05; ** p < 0.01. shRNA, short hairpin RNA

Article Snippet: An anti-MNX1 rabbit polyclonal antibody (1:500 dilution; Sigma Aldrich), an anti-CCNE1 Rabbit polyclonal antibody (1:1000 dilution;Proteintech), an anti-CCNE2 Rabbit polyclonal antibody (1:1000 dilution;Proteintech), an anti-α-tubulin mouse monoclonal antibody (1:4000 dilution; Sigma-Aldrich), an anti-Rb rabbit polyclonal antibody (1:1000 dilution; Cell Signaling Technology), an anti-p-Rb Rabbit polyclonal antibody (1:1000 dilution; Cell Signaling Technology), were used in this study.

Techniques: Activity Assay, Real-time Polymerase Chain Reaction, Expressing, Gene Expression, Plasmid Preparation, shRNA, Generated, Transformation Assay, Western Blot, Control, Luciferase, Over Expression, Knockdown, Negative Control