polyclonal rabbit anti-errα  (GeneTex)

Journal: Scientific Reports

Article Title: Long non-coding RNA SPRY4-IT1 promotes development of hepatic cellular carcinoma by interacting with ERRα and predicts poor prognosis

doi: 10.1038/s41598-017-16781-9

Figure Lengend Snippet: Knock-down of ERRα suppresses cell proliferation, colony formation, cell invasion and migration in HCC cells. ( A ) The mRNA and ( B ) protein expression of EERα in scrambled siRNA (control) or SPRY4-IT1 siRNA (siRNA) transfected HepG2 cells examined by qRT-PCR and Western blotting. ( C ) The mRNA and ( D ) protein expression of EERα in scrambled siRNA (control) or EERα siRNA (siRNA) transfected HepG2 cells examined by qRT-PCR and Western blotting. ( E ) Cell viability in scrambled siRNA (control) or EERα siRNA (siRNA) transfected HepG2 cells determined by CCK-8 assay at 48 h time point. ( F ) Colony formation assay performed in scrambled siRNA (control) or EERα siRNA (siRNA) transfected HepG2 cells. ( G ) Cell invasion and ( H ) migration assays performed in scrambled siRNA (control) or EERα siRNA (siRNA) transfected HepG2 cells. Data are shown as mean ± SD, significant differences were marked as *P < 0.05, **P < 0.01.

Article Snippet: The membranes were blocked with 5% skimmed non-fat milk for 1 h at room temperature, and then the membrane were incubated with rabbit polyclonal anti-ERRα antibody (Abcam, Cambridge, USA) at 4 °C overnight.

Techniques: Knockdown, Migration, Expressing, Control, Transfection, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Colony Assay

Journal: Scientific Reports

Article Title: Long non-coding RNA SPRY4-IT1 promotes development of hepatic cellular carcinoma by interacting with ERRα and predicts poor prognosis

doi: 10.1038/s41598-017-16781-9

Figure Lengend Snippet: ERRα overexpression antagonized the effects of SPRY4-IT1 knock-down on cell proliferation, colony formation, cell invasion and migration in HCC cells. ( A ) Cell viability in scrambled siRNA+ empty vector, SPRY4-IT1 siRNA+ empty vector, or SPRY4-IT1 siRNA+ ERRα overexpressing vector transfected HepG2 cells determined by CCK-8 assay at 48 h time point. ( B ) Colony formation assay performed in scrambled siRNA+ empty vector, SPRY4-IT1 siRNA+ empty vector, or SPRY4-IT1 siRNA+ ERRα overexpressing vector transfected HepG2 cells determined by CCK-8 assay at 48 h time point. ( C ) Cell invasion and ( D ) migration assays performed in scrambled siRNA+ empty vector, SPRY4-IT1 siRNA+ empty vector, or SPRY4-IT1 siRNA+ ERRα overexpressing vector transfected HepG2 cells. Data are shown as mean ± SD, significant differences were marked as *P < 0.05.

Article Snippet: The membranes were blocked with 5% skimmed non-fat milk for 1 h at room temperature, and then the membrane were incubated with rabbit polyclonal anti-ERRα antibody (Abcam, Cambridge, USA) at 4 °C overnight.

Techniques: Over Expression, Knockdown, Migration, Plasmid Preparation, Transfection, CCK-8 Assay, Colony Assay

Journal: PLoS ONE

Article Title: MiR-137 Targets Estrogen-Related Receptor Alpha and Impairs the Proliferative and Migratory Capacity of Breast Cancer Cells

doi: 10.1371/journal.pone.0039102

Figure Lengend Snippet: A. Schematic representation of the ERRα (ESRRA) mRNA with two putative sites (A and B) targeted by miR-137. B. Sequence alignment of predicted miR-137 target sites located within ESRRA 3′UTR showing high conservation among different species. The sequence of miR-137 target sites in ESRRA 3′UTR is shown in underlined. C. Luciferase reporter assay to verify activity of miR-137 upon the consensus miR-137 target site. HepG2 cells were transfected with Empty reporter plasmids, luciferase constructs containing perfect match miR-137 target site (miR-137 target) or mismatch miR-137 target site (△miR-137 target) and either miR-137 mimcs or NC oligos. Luciferase activity was determined 24 hr after transfection. Relative luciferase expression (firefly normalized to Renilla) values are the ratio of miR-137-treated reporter vector compared with the same NC oligos-treated reporter vector. Data are representative of at least three independent experiments. Error bars: SD. ***: P<0.0001. D. Luciferase reporter assay to evaluate the interaction between miR-137 and 3′-UTR of ESRRA. HepG2 cells were transfected with luciferase constructs containing wild-type (WT 3′UTR) or deletion mutated ESRRA 3′UTR (mutant A, mutant B and mutant C) and either miR-137 mimcs or NC oligos. Luciferase activity was determined 24 hr after transfection. Relative luciferase expression (firefly normalized to Renilla) values are the ratio of miR-137-treated reporter vector compared with the same NC oligos-treated reporter vector. Data are representative of three independent experiments. Error bars: SD. *: p<0.05, ***: P<0.0001.

Article Snippet: The antibodies used were as follows: anti-ERRα rabbit polyclonal antibody (1∶500, Millipore, cat. #07-662), anti-CyclinE1 rabbit monoclonal antibody (1∶800, Millipore cat. #04-222), anti-WNT11 rabbit polyclonal antibody (1∶600, abcam, cat.ab31962),anti-β-actin mouse monoclonal antibody (1∶500, Santa Cruz, California, USA cat. sc-47778).

Techniques: Sequencing, Luciferase, Reporter Assay, Activity Assay, Transfection, Construct, Expressing, Plasmid Preparation, Mutagenesis

Journal: PLoS ONE

Article Title: MiR-137 Targets Estrogen-Related Receptor Alpha and Impairs the Proliferative and Migratory Capacity of Breast Cancer Cells

doi: 10.1371/journal.pone.0039102

Figure Lengend Snippet: A. Western-blot analysis for ERRα protein level in normal breast epithelial cell line (MCF-10A) and five breast cancer cell lines. β-actin was used as the loading control. B. qRT-PCR analysis for miR-137 expression level. The miR-137 expression was normalized to RNU6B-small nuclear RNA. Data are representative of three independent experiments performed in triplicate. Error bars: SD.

Article Snippet: The antibodies used were as follows: anti-ERRα rabbit polyclonal antibody (1∶500, Millipore, cat. #07-662), anti-CyclinE1 rabbit monoclonal antibody (1∶800, Millipore cat. #04-222), anti-WNT11 rabbit polyclonal antibody (1∶600, abcam, cat.ab31962),anti-β-actin mouse monoclonal antibody (1∶500, Santa Cruz, California, USA cat. sc-47778).

Techniques: Western Blot, Control, Quantitative RT-PCR, Expressing

Journal: PLoS ONE

Article Title: MiR-137 Targets Estrogen-Related Receptor Alpha and Impairs the Proliferative and Migratory Capacity of Breast Cancer Cells

doi: 10.1371/journal.pone.0039102

Figure Lengend Snippet: A. Western blot analysis for ERRα protein level and qRT-PCR analysis for ERRα mRNA level in SK-BR-3 cells 48 hr after transfection regent treatment (mock) or transfection with indicated RNA oligonucleotides (50 nM). B. Western blot analysis for ERRα protein level and qRT-PCR analysis for ERRα mRNA level in SK-BR-3 cells 48 hr after transfection regent treatment (mock) or cotransfection with equal amount of indicated RNA oligonucleotides. ERRα mRNA expression was normalized to β-actin mRNA expression. The relative level of ERRα expression determined using the 2- △△CT method. Data are representative of three independent experiments performed in triplicate. Error bars: SD; *: p<0.05; ***: P<0.0001.

Article Snippet: The antibodies used were as follows: anti-ERRα rabbit polyclonal antibody (1∶500, Millipore, cat. #07-662), anti-CyclinE1 rabbit monoclonal antibody (1∶800, Millipore cat. #04-222), anti-WNT11 rabbit polyclonal antibody (1∶600, abcam, cat.ab31962),anti-β-actin mouse monoclonal antibody (1∶500, Santa Cruz, California, USA cat. sc-47778).

Techniques: Western Blot, Quantitative RT-PCR, Transfection, Cotransfection, Expressing

Journal: PLoS ONE

Article Title: MiR-137 Targets Estrogen-Related Receptor Alpha and Impairs the Proliferative and Migratory Capacity of Breast Cancer Cells

doi: 10.1371/journal.pone.0039102

Figure Lengend Snippet: Breast cancer cell lines (MCF-7, BT-474, SK-BR-3 and MDA-MB-231) were transfected with 50 nM NC oligos, si-ERRα or miR-137 mimics or treated with transfection reagent alone (mock) and seeded in 96-well plates. Plates were harvested at 1, 2, 3, 4, 5 days after seeding for CCK-8 assay.

Article Snippet: The antibodies used were as follows: anti-ERRα rabbit polyclonal antibody (1∶500, Millipore, cat. #07-662), anti-CyclinE1 rabbit monoclonal antibody (1∶800, Millipore cat. #04-222), anti-WNT11 rabbit polyclonal antibody (1∶600, abcam, cat.ab31962),anti-β-actin mouse monoclonal antibody (1∶500, Santa Cruz, California, USA cat. sc-47778).

Techniques: Transfection, CCK-8 Assay

Journal: PLoS ONE

Article Title: MiR-137 Targets Estrogen-Related Receptor Alpha and Impairs the Proliferative and Migratory Capacity of Breast Cancer Cells

doi: 10.1371/journal.pone.0039102

Figure Lengend Snippet: A. Western blot analysis for ERRα and CylinE1 protein level and qRT-PCR analysis for CCNE1 mRNA level in SK-BR-3 cells 48 hr after DMSO or XCT-790 treatment. B. Western blot analysis for CylinE1 protein level and qRT-PCR analysis for CCNE1 mRNA level in SK-BR-3 cells 48 hr after transfection regent treatment (mock) or transfection with NC oligos or si-ERRα. C. Western blot analysis for CylinE1 protein level and qRT-PCR analysis for CCNE1 mRNA level in SK-BR-3 48 hr after transfection regent treatment (mock) or co-transfection with equal amount of indicated RNA oligonucleotides. CCNE1 mRNA expression was normalized to β-actin mRNA expression. The relative level of CCNE1 mRNA was determined using the 2- △△CT method. Data are representative of three independent experiments performed in triplicate. Error bars: SD; *: p<0.05; **: P<0.01; ***: P<0.0001.

Article Snippet: The antibodies used were as follows: anti-ERRα rabbit polyclonal antibody (1∶500, Millipore, cat. #07-662), anti-CyclinE1 rabbit monoclonal antibody (1∶800, Millipore cat. #04-222), anti-WNT11 rabbit polyclonal antibody (1∶600, abcam, cat.ab31962),anti-β-actin mouse monoclonal antibody (1∶500, Santa Cruz, California, USA cat. sc-47778).

Techniques: Western Blot, Quantitative RT-PCR, Transfection, Cotransfection, Expressing

Journal: PLoS ONE

Article Title: MiR-137 Targets Estrogen-Related Receptor Alpha and Impairs the Proliferative and Migratory Capacity of Breast Cancer Cells

doi: 10.1371/journal.pone.0039102

Figure Lengend Snippet: A. re-expression of ERRα (without 3′-UTR) in SK-BR-3 cells transfected with miR-137 reversed the decrease of CCNE1 expression induced by miR-137. qRT-PCR analysis for ERRα and CCNE1 mRNA level and western blot analysis for ERRα and CylinE1 protein level in SK-BR-3 cells 48 hr after transfection. CCNE1 or ERRα mRNA expression was normalized to β-actin mRNA expression. The relative level of CCNE1 or ERRα determined using the 2- △△CT method. Data are representative of three independent experiments performed in duplicate. Error bars: SD; **: P<0.01; ***: P<0.0001. B. re-expression of ERRα (without 3′-UTR) in SK-BR-3 cells transfected with miR-137 partly rescued the arrested cell cycle progression. Cell cycle analysis using propidium iodide staining and flow cytometry was performed in SK-BR-3 cells transfected with indicated RNA oligonucleotides (50 nM) and plasmids (300 ng) for three days. The percentage of cells in each cell cycle phases was quantified. Error bars: SD; *: p<0.05; ***: P<0.0001. C. re-expression of ERRα (without 3′-UTR) in SK-BR-3 cells transfected with miR-137 partly rescued the impaired proliferation capacity. SK-BR-3 cells were transfected with indicated RNA oligonucleotides (50 nM) and plasmids (30ng) and seeded in 96-well plates. Plates were harvested at 1, 3, 5, 7 days after seeding. Cell numbers were determined by CCK-8 assay.

Article Snippet: The antibodies used were as follows: anti-ERRα rabbit polyclonal antibody (1∶500, Millipore, cat. #07-662), anti-CyclinE1 rabbit monoclonal antibody (1∶800, Millipore cat. #04-222), anti-WNT11 rabbit polyclonal antibody (1∶600, abcam, cat.ab31962),anti-β-actin mouse monoclonal antibody (1∶500, Santa Cruz, California, USA cat. sc-47778).

Techniques: Expressing, Transfection, Quantitative RT-PCR, Western Blot, Cell Cycle Assay, Staining, Flow Cytometry, CCK-8 Assay

Journal: PLoS ONE

Article Title: MiR-137 Targets Estrogen-Related Receptor Alpha and Impairs the Proliferative and Migratory Capacity of Breast Cancer Cells

doi: 10.1371/journal.pone.0039102

Figure Lengend Snippet: A. re-expression of ERRα (without 3′-UTR) in MDA-MB-231 cells restored the impaired migratory capacity induced by miR-137. MDA-MB-231 cells were co-transfected with indicated RNA oligonucleotides (50 nM) and plasmids (1 µg), and serum starved for 12 hr, followed by assessment of cell invasion and viability. Error bars: SD; *: p<0.05; **: P<0.01; ***: P<0.0001. B. re-expression of ERRα (without 3′-UTR) in MDA-MB-231 cells reversed the decrease of WNT11 expression induced by miR-137. MDA-MB-231 cells were co-transfected with indicated RNA oligonucleotides (50 nM) and plasmids (1 µg). 48 hr after transfection, protein and mRNA levels of WNT11 and ERRα were assayed using western bolt and qRT-PCR respectively. WNT11 or ERRα mRNA expression was normalized to β-actin mRNA expression. The relative level of WNT11 or ERRα determined using the 2- △△CT method. Data are representative of three independent experiments performed in duplicate. Error bars: SD; **: P<0.01; ***: P<0.0001.

Article Snippet: The antibodies used were as follows: anti-ERRα rabbit polyclonal antibody (1∶500, Millipore, cat. #07-662), anti-CyclinE1 rabbit monoclonal antibody (1∶800, Millipore cat. #04-222), anti-WNT11 rabbit polyclonal antibody (1∶600, abcam, cat.ab31962),anti-β-actin mouse monoclonal antibody (1∶500, Santa Cruz, California, USA cat. sc-47778).

Techniques: Expressing, Transfection, Western Blot, Quantitative RT-PCR