primary antibody against e2f3 sc-878  (Santa Cruz Biotechnology)

Journal: Molecular Medicine Reports

Article Title: miR-34a suppresses proliferation and induces apoptosis of human lens epithelial cells by targeting E2F3

doi: 10.3892/mmr.2016.5901

Figure Lengend Snippet: E2F3 was a direct target of miR-34a. (A and B) Schematic representation of WT and mut putative miR-34a-binding sites in the 3′-UTR of E2F3. (C) The 293T cells were transfected with miR-34a mimic and psiCHECK2 containing either the WT or mut putative binding sites for miR-34a, and 48 h later the luciferase assay was performed. The data indicated that miR-34a mimic reduced the Renilla activity in the reporter construct containing the E2F3 site (1.87±0.13), whereas no effect was observed with a construct containing a mut E2F3 seed site (3.46±0.09) and mimic control (3.27±0.04). Data are presented as the mean ± standard deviation; *P<0.05. miR, microRNA; WT, wild-type; mut, mutant; UTR, untranslated region; NS, non-significant.

Article Snippet: Subsequent to blocking in Tris-buffered saline with Tween-20 (TBST) containing 25 mmol/l Tris-HCl, pH 7.5, 137 mmol/l NaCl, 2.7 mmol/l KCl and 0.05% Tween-20 with 5% nonfat milk for 1 h at 37°C, the membranes were incubated with the primary antibody against E2F3 (sc-878; 1:200; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) or GAPDH (ab9485; 1:2,500; Abcam, Cambridge, MA, USA) in TBST with 5% nonfat milk at 4°C overnight.

Techniques: Binding Assay, Transfection, Luciferase, Activity Assay, Construct, Control, Standard Deviation, Mutagenesis

Journal: Molecular Medicine Reports

Article Title: miR-34a suppresses proliferation and induces apoptosis of human lens epithelial cells by targeting E2F3

doi: 10.3892/mmr.2016.5901

Figure Lengend Snippet: miR-34a and siE2F3 reduce E2F3 expression. SRA01/04 cells transfected with miR-34a mimic or siE2F3 exhibited reduced E2F3 expression compared with the mock and mimic control. Data are presented as the mean ± standard deviation; *P<0.05. miR, microRNA; si, small interfering; NS, non-significant.

Article Snippet: Subsequent to blocking in Tris-buffered saline with Tween-20 (TBST) containing 25 mmol/l Tris-HCl, pH 7.5, 137 mmol/l NaCl, 2.7 mmol/l KCl and 0.05% Tween-20 with 5% nonfat milk for 1 h at 37°C, the membranes were incubated with the primary antibody against E2F3 (sc-878; 1:200; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) or GAPDH (ab9485; 1:2,500; Abcam, Cambridge, MA, USA) in TBST with 5% nonfat milk at 4°C overnight.

Techniques: Expressing, Transfection, Control, Standard Deviation

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Bromodomain and extraterminal protein inhibitor JQ1 suppresses thyroid tumor growth in a mouse model

doi: 10.1158/1078-0432.CCR-16-0914

Figure Lengend Snippet: JQ1 alters cyclin D1-CDK4-Rb-E2F3 signaling and AT2A/HEXIM1 abundance in thyroid tumors of ThrbPV/PVKrasG12D mice. (A-I) Total protein extracts were prepared from thyroid tumors. Western blot analysis was carried out for p-Rb(S780) (a), total Rb (b), E3F3 (c), CDK4 (d), cyclin D1 (e), KAT2A (f), HEXIM1 (g), and GAPDH (h), as described in Materials and Methods. (A-II) Quantitative analysis of relative protein expression levels of the ratio of p-Rb(S780) to total Rb (a), E3F3 (b), CDK4 (c), cyclin D1 (d), KAT2A (e), HEXIM1 (f) in mice treated with vehicle or JQ1. (B). Decreased Kat2a (panel a) and increased Hexim1 (panel b) mRNA expression in thyroid tumors of ThrbPV/PVKrasG12D mice treated with JQ1. Total RNAs prepared from thyroid tumors of the ThrbPV/PVKrasG12D mice treated with vehicle or JQ1 were used for the analysis of RNA expression.

Article Snippet: The E2F3 primary antibody (sc-878) and Rb (sc-50) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Western Blot, Expressing, RNA Expression