polyclonal rabbit anti yap1 primary antibody  (Proteintech)

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

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

Journal: Cancer discovery

Article Title: Bi-allelic Alteration and Dysregulation of the Hippo Pathway in Mucinous Tubular and Spindle Cell Carcinoma of the Kidney

doi: 10.1158/2159-8290.CD-16-0267

Figure Lengend Snippet: Hippo pathway gene alterations in MTSCC. A. Schematic representation of the Hippo pathway gene aberrations discovered in MTSCC. PFAM domains and scale depicting amino acid numbers are presented for each protein schema. Indels (red dots), missense SNVs (green dots), nonsense (black dots) and splice site SNVs (yellow dots). B. An example of YAP1 nuclear expression in MTSCC case RC_1100 as revealed by immunohistochemistry. Benign background renal parenchyma (left panel, low power view 100× with scale bar 200 microns, and inset 400×) demonstrates patchy and weak predominantly nuclear YAP1 staining (with focal cytoplasmic expression). Tumor areas of MTSCC (right panel, low power view 100× with scale bar 200 microns, and inset 400×) from the same case demonstrate moderate to strong nuclear YAP1 expression (with focal cytoplasmic staining). C. MTSCC Hippo-pathway mutations in the cohort (top panel; red indels, yellow- splice site, black-nonsense, green- missense, cells crossed white line indicates bi-alleic loss) represented along with case wise evaluation of YAP1 protein nuclear localization (bottom panel; dark brown- present; white- absent) and increased YAP1 protein expression in tumor compared to background benign renal parenchyma (bottom panel; light brown- present; white- absent). D. Hippo signaling pathway schematic with alterations identified in this study highlighted. Pathway members with mutations are indicated (filled boxes) and numbers within represent the cases with that particular somatic aberration (except in the nuclear YAP1 box where the numbers represent immunohistochemistry staining data for cases with nuclear localization); putative tumor suppressors are colored green while oncogenes are colored red.

Article Snippet: In the current study, IHC were performed on representative whole tumor sections from the MTSCC cases with anti-YAP1 primary antibody (Cell Signaling Technology: / Danvers, MA / 01923 YAP1 Rabbit polyclonal cat# 4912s; 1 in 300 dilution).

Techniques: Expressing, Immunohistochemistry, Staining