Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: Upregulated microRNA let-7a accelerates apoptosis and inhibits proliferation in uterine junctional zone smooth muscle cells in adenomyosis under conditions of a normal activated hippo-YAP1 axis

doi: 10.1186/s12958-021-00753-w

Figure Lengend Snippet: The expression levels of let-7a, YAP1 and TAZ in JZ SMCs in the ADS group and control group. a The mRNA expression level of let-7a determined by RT-qPCR and normalized against that of U6. b The mRNA expression level of YAP1 determined by RT-qPCR and normalized against that of β-actin. c The mRNA expression level of TAZ determined by RT-qPCR and normalized against that of β-actin. d The protein expression levels of YAP1 and TAZ determined by western blotting and normalized against that of α-tubulin. e The relative fold protein expression of YAP1 normalized against α-tubulin. f The relative fold protein expression of TAZ normalized against α-tubulin. These experiments were performed two times with three replicates in each experiment. Significance was determined by Student’s t test; ** p < 0.01, * p < 0.05

Article Snippet: Next, the membranes were washed and incubated with an anti-YAP1 rabbit monoclonal antibody (Beyotime Institute of Biotechnology, China), an anti-phospho-YAP1 rabbit monoclonal antibody (1:1000; Beyotime Institute of Biotechnology, China), an anti-TAZ rabbit polyclonal antibody (1:1000, Beyotime Institute of Biotechnology, China), and an anti-α-tubulin rabbit monoclonal antibody (1:1000, Cell Signalling Technology) overnight at 4 °C with gentle agitation.

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

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: Upregulated microRNA let-7a accelerates apoptosis and inhibits proliferation in uterine junctional zone smooth muscle cells in adenomyosis under conditions of a normal activated hippo-YAP1 axis

doi: 10.1186/s12958-021-00753-w

Figure Lengend Snippet: The expression levels of YAP1, TAZ and phospho-YAP1 in JZ SMCs in the ADS group. a Cells were transfected with the let-7a overexpression lentiviral vector GV280 or lentiviral null vector for 72 h and then used for western blotting. b The relative fold protein expression of YAP1 normalized against α-tubulin. c The relative fold protein expression of TAZ normalized against α-tubulin. d The relative fold protein expression of phospho-YAP1 normalized against α-tubulin. These experiments were performed two times with three replicates in each experiment. Significance was determined by Student’s t test; **** p < 0.0001, *** p < 0.001

Article Snippet: Next, the membranes were washed and incubated with an anti-YAP1 rabbit monoclonal antibody (Beyotime Institute of Biotechnology, China), an anti-phospho-YAP1 rabbit monoclonal antibody (1:1000; Beyotime Institute of Biotechnology, China), an anti-TAZ rabbit polyclonal antibody (1:1000, Beyotime Institute of Biotechnology, China), and an anti-α-tubulin rabbit monoclonal antibody (1:1000, Cell Signalling Technology) overnight at 4 °C with gentle agitation.

Techniques: Expressing, Transfection, Over Expression, Plasmid Preparation, Western Blot

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: Upregulated microRNA let-7a accelerates apoptosis and inhibits proliferation in uterine junctional zone smooth muscle cells in adenomyosis under conditions of a normal activated hippo-YAP1 axis

doi: 10.1186/s12958-021-00753-w

Figure Lengend Snippet: The expression levels of YAP, TAZ and phospho-YAP1 in JZ SMCs in the verteporfin-ADS group and ADS group and CCK-8 assay results for the verteporfin-ADS group and negative control group. a Cells were co-cultured with 1 μM verteporfin and then used for western blotting. b The relative fold protein expression of YAP1 normalized against α-tubulin. c The relative fold protein expression of TAZ normalized against α-tubulin. d The relative fold protein expression of phospho-YAP1 normalized against α-tubulin. e Cells were treated with verteporfin, and then a CCK-8 assay was used to test the proliferative ability of ADS JZ SMCs. These experiments were performed two times with three replicates in each experiment. Significance was determined by Student’s t test; **** p < 0.0001, ** p < 0.01, * p < 0.05

Article Snippet: Next, the membranes were washed and incubated with an anti-YAP1 rabbit monoclonal antibody (Beyotime Institute of Biotechnology, China), an anti-phospho-YAP1 rabbit monoclonal antibody (1:1000; Beyotime Institute of Biotechnology, China), an anti-TAZ rabbit polyclonal antibody (1:1000, Beyotime Institute of Biotechnology, China), and an anti-α-tubulin rabbit monoclonal antibody (1:1000, Cell Signalling Technology) overnight at 4 °C with gentle agitation.

Techniques: Expressing, CCK-8 Assay, Negative Control, Cell Culture, Western Blot