polyclonal rabbit anti phospho yap1 primary antibody  (Cell Signaling Technology Inc)

Journal: Experimental and Therapeutic Medicine

Article Title: gga-microRNA-375 negatively regulates the cell cycle and proliferation by targeting Yes-associated protein 1 in DF-1 cells

doi: 10.3892/etm.2020.8711

Figure Lengend Snippet: RNA oligoribonucleotides used for cell transfection.

Article Snippet: Membranes with corresponding proteins were incubated with the polyclonal rabbit anti-YAP1 primary antibody (cat. no. 13584-1-AP; 1:1,000; ProteinTech Group, Inc.), polyclonal rabbit anti-phospho-YAP1 primary antibody (cat. no. 13008; 1:11,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-cyclin D1 primary antibody (cat. no. 55506; 1:1,000; Cell Signaling Technology, Inc.), polyclonal mouse anti-p53 primary antibody (cat. no. 2524; 1:11,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-cyclin E primary antibody (cat. no. 4129; 1:1,000; Cell Signaling Technology, Inc.), monoclonal mouse anti-p27 primary antibody (the antibody was generated and generously provided by Professor Wencheng Lin; 1:1,000), polyclonal rabbit anti-MST1 primary antibody (cat. no. 3682; 1:1,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-SAV1 primary antibody (cat. no. 3507; 1:1,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-MOB1 primary antibody (cat. no. 3863; 1:1,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-LATS1 primary antibody (cat. no. 9153; 1:1,000; Cell Signaling Technology, Inc.), monoclonal rabbit anti-GAPDH primary antibody (cat. no. 2118; 1:1,000; Cell Signaling Technology, Inc.) and polyclonal mouse anti-β-actin primary antibody (cat. no. 33308; 1:1,000; Bioss Antibodies, Inc.) overnight at 4 ̊C.

Techniques: Transfection, Sequencing

Journal: Experimental and Therapeutic Medicine

Article Title: gga-microRNA-375 negatively regulates the cell cycle and proliferation by targeting Yes-associated protein 1 in DF-1 cells

doi: 10.3892/etm.2020.8711

Figure Lengend Snippet: Primers used for reverse transcription-quantitative PCR.

Article Snippet: Membranes with corresponding proteins were incubated with the polyclonal rabbit anti-YAP1 primary antibody (cat. no. 13584-1-AP; 1:1,000; ProteinTech Group, Inc.), polyclonal rabbit anti-phospho-YAP1 primary antibody (cat. no. 13008; 1:11,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-cyclin D1 primary antibody (cat. no. 55506; 1:1,000; Cell Signaling Technology, Inc.), polyclonal mouse anti-p53 primary antibody (cat. no. 2524; 1:11,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-cyclin E primary antibody (cat. no. 4129; 1:1,000; Cell Signaling Technology, Inc.), monoclonal mouse anti-p27 primary antibody (the antibody was generated and generously provided by Professor Wencheng Lin; 1:1,000), polyclonal rabbit anti-MST1 primary antibody (cat. no. 3682; 1:1,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-SAV1 primary antibody (cat. no. 3507; 1:1,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-MOB1 primary antibody (cat. no. 3863; 1:1,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-LATS1 primary antibody (cat. no. 9153; 1:1,000; Cell Signaling Technology, Inc.), monoclonal rabbit anti-GAPDH primary antibody (cat. no. 2118; 1:1,000; Cell Signaling Technology, Inc.) and polyclonal mouse anti-β-actin primary antibody (cat. no. 33308; 1:1,000; Bioss Antibodies, Inc.) overnight at 4 ̊C.

Techniques:

Journal: Experimental and Therapeutic Medicine

Article Title: gga-microRNA-375 negatively regulates the cell cycle and proliferation by targeting Yes-associated protein 1 in DF-1 cells

doi: 10.3892/etm.2020.8711

Figure Lengend Snippet: Overexpression of gga-miR-375 inhibits proliferation and the cell cycle in DF-1 cells. DF-1 cells transfected with gga-miR-375 mimic decreases YAP1 (A) protein and (B) mRNA expression levels after 48 h of transfection. gga-miR-NC and mock are the control groups. (C) Overexpression of gga-miR-375 inhibits cell proliferation as indicated by Cell Counting Kit-8 assay results. (D) Overexpression of gga-miR-375 inhibits cell cycle progression as measured with a FACSCalibur flow cytometer. (E) Overexpression of gga-miR-375 downregulates the expression levels of YAP1, cyclin D1 and cyclin E. Data are presented as the mean ± SD of three independent experiments. * * P<0.01. NC, negative control; miR, microRNA; YAP1, Yes-associated protein 1; OD, optical density; PI, propidium iodide.

Article Snippet: Membranes with corresponding proteins were incubated with the polyclonal rabbit anti-YAP1 primary antibody (cat. no. 13584-1-AP; 1:1,000; ProteinTech Group, Inc.), polyclonal rabbit anti-phospho-YAP1 primary antibody (cat. no. 13008; 1:11,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-cyclin D1 primary antibody (cat. no. 55506; 1:1,000; Cell Signaling Technology, Inc.), polyclonal mouse anti-p53 primary antibody (cat. no. 2524; 1:11,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-cyclin E primary antibody (cat. no. 4129; 1:1,000; Cell Signaling Technology, Inc.), monoclonal mouse anti-p27 primary antibody (the antibody was generated and generously provided by Professor Wencheng Lin; 1:1,000), polyclonal rabbit anti-MST1 primary antibody (cat. no. 3682; 1:1,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-SAV1 primary antibody (cat. no. 3507; 1:1,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-MOB1 primary antibody (cat. no. 3863; 1:1,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-LATS1 primary antibody (cat. no. 9153; 1:1,000; Cell Signaling Technology, Inc.), monoclonal rabbit anti-GAPDH primary antibody (cat. no. 2118; 1:1,000; Cell Signaling Technology, Inc.) and polyclonal mouse anti-β-actin primary antibody (cat. no. 33308; 1:1,000; Bioss Antibodies, Inc.) overnight at 4 ̊C.

Techniques: Over Expression, Transfection, Expressing, Control, Cell Counting, Flow Cytometry, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: gga-microRNA-375 negatively regulates the cell cycle and proliferation by targeting Yes-associated protein 1 in DF-1 cells

doi: 10.3892/etm.2020.8711

Figure Lengend Snippet: Knockdown of gga-miR-375 promotes proliferation and the cell cycle in DF-1 cells. (A) DF-1 cells transfected with anti-gga-miR-375 had decreased gga-miR-375 mRNA expression at 48 h following transcription. Anti-gga-miR-con group and mock group were used as the controls. DF-1 cells transfected with anti-gga-miR-375 exhibited increased (B) YAP1 protein and (C) mRNA expression levels compared with the control groups. (D) Overexpression of anti-gga-miR-375 promotes cell proliferation as indicated by Cell Counting Kit-8 assay results 24, 48 and 72 h after transfection. (E) Overexpression of anti-gga-miR-375 promotes the cell cycle as demonstrated by flow cytometry results. (F) DF-1 cells transfected with anti-gga-miR-375 had upregulated expression levels of YAP1, cyclin D1 and cyclin E. Anti-gga-miR-con and mock groups were used as the control groups. Data are presented as the mean ± SD of three independent experiments. * * P<0.01 vs. anti-miR-con group. miR, microRNA; YAP1, Yes-associated protein 1; NC, negative control; con, control; OD, optical density.

Article Snippet: Membranes with corresponding proteins were incubated with the polyclonal rabbit anti-YAP1 primary antibody (cat. no. 13584-1-AP; 1:1,000; ProteinTech Group, Inc.), polyclonal rabbit anti-phospho-YAP1 primary antibody (cat. no. 13008; 1:11,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-cyclin D1 primary antibody (cat. no. 55506; 1:1,000; Cell Signaling Technology, Inc.), polyclonal mouse anti-p53 primary antibody (cat. no. 2524; 1:11,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-cyclin E primary antibody (cat. no. 4129; 1:1,000; Cell Signaling Technology, Inc.), monoclonal mouse anti-p27 primary antibody (the antibody was generated and generously provided by Professor Wencheng Lin; 1:1,000), polyclonal rabbit anti-MST1 primary antibody (cat. no. 3682; 1:1,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-SAV1 primary antibody (cat. no. 3507; 1:1,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-MOB1 primary antibody (cat. no. 3863; 1:1,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-LATS1 primary antibody (cat. no. 9153; 1:1,000; Cell Signaling Technology, Inc.), monoclonal rabbit anti-GAPDH primary antibody (cat. no. 2118; 1:1,000; Cell Signaling Technology, Inc.) and polyclonal mouse anti-β-actin primary antibody (cat. no. 33308; 1:1,000; Bioss Antibodies, Inc.) overnight at 4 ̊C.

Techniques: Knockdown, Transfection, Expressing, Control, Over Expression, Cell Counting, Flow Cytometry, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: gga-microRNA-375 negatively regulates the cell cycle and proliferation by targeting Yes-associated protein 1 in DF-1 cells

doi: 10.3892/etm.2020.8711

Figure Lengend Snippet: OE of YAP1 promotes proliferation and cell cycle in DF-1 cells. (A) DF-1 cells transfected with pRK5-Flag-YAP1 were harvested after 48 h for western blot analysis with antibodies against YAP1 and β-actin. (B) OE of YAP1 promotes cell proliferation as identified by Cell Counting Kit-8 assay results 24, 48 and 72 h after transfection. (C) OE of YAP1 promotes the cell cycle, as demonstrated by flow cytometry. (D) OE of YAP1 promotes the protein expression levels of cyclin D1 and cyclin E after 48 h of transfection. Data are presented as the mean ± SD of three independent experiments. * * P<0.01 vs. EV group. EV, empty vector; YAP1, Yes-associated protein 1; OD, optical density; OE, overexpression; PI, propidium iodide.

Article Snippet: Membranes with corresponding proteins were incubated with the polyclonal rabbit anti-YAP1 primary antibody (cat. no. 13584-1-AP; 1:1,000; ProteinTech Group, Inc.), polyclonal rabbit anti-phospho-YAP1 primary antibody (cat. no. 13008; 1:11,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-cyclin D1 primary antibody (cat. no. 55506; 1:1,000; Cell Signaling Technology, Inc.), polyclonal mouse anti-p53 primary antibody (cat. no. 2524; 1:11,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-cyclin E primary antibody (cat. no. 4129; 1:1,000; Cell Signaling Technology, Inc.), monoclonal mouse anti-p27 primary antibody (the antibody was generated and generously provided by Professor Wencheng Lin; 1:1,000), polyclonal rabbit anti-MST1 primary antibody (cat. no. 3682; 1:1,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-SAV1 primary antibody (cat. no. 3507; 1:1,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-MOB1 primary antibody (cat. no. 3863; 1:1,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-LATS1 primary antibody (cat. no. 9153; 1:1,000; Cell Signaling Technology, Inc.), monoclonal rabbit anti-GAPDH primary antibody (cat. no. 2118; 1:1,000; Cell Signaling Technology, Inc.) and polyclonal mouse anti-β-actin primary antibody (cat. no. 33308; 1:1,000; Bioss Antibodies, Inc.) overnight at 4 ̊C.

Techniques: Transfection, Western Blot, Cell Counting, Flow Cytometry, Expressing, Plasmid Preparation, Over Expression

Journal: Experimental and Therapeutic Medicine

Article Title: gga-microRNA-375 negatively regulates the cell cycle and proliferation by targeting Yes-associated protein 1 in DF-1 cells

doi: 10.3892/etm.2020.8711

Figure Lengend Snippet: Knockdown of YAP1 inhibits proliferation and the cell cycle in DF-1 cells. (A) DF-1 cells were transfected with siRNA-YAP1 and harvested 48 h later for western blot analysis with antibodies against YAP1 and β-actin. (B) Knockdown of YAP1 inhibits cell proliferation as demonstrated by Cell Counting Kit-8 assay results 24, 48 and 72 h after transfection. (C) Flow cytometry results indicated that knockdown of YAP1 inhibited the cell cycle. (D) Knockdown of YAP1 inhibits the protein expression levels of cyclin D1 and cyclin E after 48 h transfection. Data are presented as the mean ± SD of three independent experiments. * * P<0.01 vs. siRNA-Scra group. YAP1, Yes-associated protein 1; siRNA, small interfering RNA; Scra, scramble; PI, propidium iodide; OD, optical density.

Article Snippet: Membranes with corresponding proteins were incubated with the polyclonal rabbit anti-YAP1 primary antibody (cat. no. 13584-1-AP; 1:1,000; ProteinTech Group, Inc.), polyclonal rabbit anti-phospho-YAP1 primary antibody (cat. no. 13008; 1:11,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-cyclin D1 primary antibody (cat. no. 55506; 1:1,000; Cell Signaling Technology, Inc.), polyclonal mouse anti-p53 primary antibody (cat. no. 2524; 1:11,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-cyclin E primary antibody (cat. no. 4129; 1:1,000; Cell Signaling Technology, Inc.), monoclonal mouse anti-p27 primary antibody (the antibody was generated and generously provided by Professor Wencheng Lin; 1:1,000), polyclonal rabbit anti-MST1 primary antibody (cat. no. 3682; 1:1,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-SAV1 primary antibody (cat. no. 3507; 1:1,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-MOB1 primary antibody (cat. no. 3863; 1:1,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-LATS1 primary antibody (cat. no. 9153; 1:1,000; Cell Signaling Technology, Inc.), monoclonal rabbit anti-GAPDH primary antibody (cat. no. 2118; 1:1,000; Cell Signaling Technology, Inc.) and polyclonal mouse anti-β-actin primary antibody (cat. no. 33308; 1:1,000; Bioss Antibodies, Inc.) overnight at 4 ̊C.

Techniques: Knockdown, Transfection, Western Blot, Cell Counting, Flow Cytometry, Expressing, Small Interfering RNA

Journal: Experimental and Therapeutic Medicine

Article Title: gga-microRNA-375 negatively regulates the cell cycle and proliferation by targeting Yes-associated protein 1 in DF-1 cells

doi: 10.3892/etm.2020.8711

Figure Lengend Snippet: Knockdown of gga-miR-375 expression promotes the cell cycle and cell proliferation by targeting YAP1. (A) DF-1 cells transfected with anti-gga-miR-375 had decreased gga-miR-375 mRNA expression at 48 h as assessed by reverse transcription-quantitative PCR. Anti-gga-miR-con and mock groups are the controls. (B) Protein expression of YAP1 was measure after DF-1 cells were transfected for 48 h with anti-gga-miR-375. (C) Protein expression levels of YAP1 after 48 h transfection with anti-gga-miR-375 or co-transfected with anti-gga-miR-375 + siRNA-YAP1. (D) Knockdown of gga-miR-375 promoted cell proliferation by targeting YAP1, as identified by Cell Counting Kit-8 assay results after 24, 48 and 72 h. (E) Flow cytometry results indicated that knockdown of gga-miR-375 expression promoted the cell cycle by targeting YAP1. The cell cycle assay was performed after knockdown of gga-miR-375 or gga-miR-375 + YAP1 in DF-1 cells and evaluated with a FACSCalibur flow cytometer 48 h after transfection. (F) DF-1 cells transfected with anti-gga-miR-375 or anti-gga-miR-375 + siRNA-YAP1 were harvested 48 h after transfection for western blot analysis with antibodies against YAP1, cyclin D1, cyclin E and β-actin. Data are presented as the mean ± SD of three independent experiments. * * P<0.01. YAP1, yes-associated protein 1; miR, microRNA; siRNA, small interfering RNA; PI, propidium iodide; OD, optical density; Scra, scramble.

Article Snippet: Membranes with corresponding proteins were incubated with the polyclonal rabbit anti-YAP1 primary antibody (cat. no. 13584-1-AP; 1:1,000; ProteinTech Group, Inc.), polyclonal rabbit anti-phospho-YAP1 primary antibody (cat. no. 13008; 1:11,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-cyclin D1 primary antibody (cat. no. 55506; 1:1,000; Cell Signaling Technology, Inc.), polyclonal mouse anti-p53 primary antibody (cat. no. 2524; 1:11,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-cyclin E primary antibody (cat. no. 4129; 1:1,000; Cell Signaling Technology, Inc.), monoclonal mouse anti-p27 primary antibody (the antibody was generated and generously provided by Professor Wencheng Lin; 1:1,000), polyclonal rabbit anti-MST1 primary antibody (cat. no. 3682; 1:1,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-SAV1 primary antibody (cat. no. 3507; 1:1,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-MOB1 primary antibody (cat. no. 3863; 1:1,000; Cell Signaling Technology, Inc.), polyclonal rabbit anti-LATS1 primary antibody (cat. no. 9153; 1:1,000; Cell Signaling Technology, Inc.), monoclonal rabbit anti-GAPDH primary antibody (cat. no. 2118; 1:1,000; Cell Signaling Technology, Inc.) and polyclonal mouse anti-β-actin primary antibody (cat. no. 33308; 1:1,000; Bioss Antibodies, Inc.) overnight at 4 ̊C.

Techniques: Knockdown, Expressing, Transfection, Reverse Transcription, Real-time Polymerase Chain Reaction, Cell Counting, Flow Cytometry, Cell Cycle Assay, Western Blot, Small Interfering RNA