Journal: British journal of pharmacology

Article Title: The orphan receptor NOR1 participates in isoprenaline-induced cardiac hypertrophy by regulating PARP-1.

doi: 10.1111/bph.13091

Figure Lengend Snippet: Figure 1 NOR1 expression was significantly increased in NRCMs and heart tissues following isoprenaline treatment. (A and B) Primary NRCMs were treated with 1 μM isoprenaline (ISO) for indicated time points. The mRNA and protein levels of NOR1 were detected by qRT-PCR and Western blot respectively. (C and D) SD rats were subjected to s.c. injection of 1.5 mg·kg−1·d−1 isoprenaline or normal saline (NS) for 7 d, and then the NOR1 expression was measured. The results were normalized by GAPDH for mRNA or Lamin for protein expression. Data were presented as means ± SE. *P < 0.05 versus control or NS group, n = 5 independent experiments.

Article Snippet: For immunoprecipitation, rabbit anti-NOR1 polyclonal antibody (diluted 1:10) and mouse anti-GFP monoclonal antibody (diluted 1:10) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

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

Journal: British journal of pharmacology

Article Title: The orphan receptor NOR1 participates in isoprenaline-induced cardiac hypertrophy by regulating PARP-1.

doi: 10.1111/bph.13091

Figure Lengend Snippet: Figure 2 Knockdown of NOR1 blocked the hypertrophic responses induced by isoprenaline (ISO). (A and B) Cultured NRCMs were transfected with three different siRNA for NOR1 (S1, S3, S3) or negative control (NC) for 24 h, and the silencing efficacy was measured by qRT-PCR and Western blot. Cells were transfected with NOR1 siRNA (siNOR1) before incubation with 1 μM isoprenaline for 24 h. (C) Expression of NOR1 protein was detected by Western blot. (D) ANF and BNP mRNA levels were determined by qRT-PCR. (E) Cell surface area was measured by rhodamine- phalloidin staining. Data were presented as means ± SE. *P < 0.05, versus NC group, #P < 0.05 versus NC treated with isoprenaline, n = 5 independent experiments.

Article Snippet: For immunoprecipitation, rabbit anti-NOR1 polyclonal antibody (diluted 1:10) and mouse anti-GFP monoclonal antibody (diluted 1:10) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

Techniques: Knockdown, Cell Culture, Transfection, Negative Control, Quantitative RT-PCR, Western Blot, Incubation, Expressing, Staining

Journal: British journal of pharmacology

Article Title: The orphan receptor NOR1 participates in isoprenaline-induced cardiac hypertrophy by regulating PARP-1.

doi: 10.1111/bph.13091

Figure Lengend Snippet: Figure 3 NOR1 overexpression led to hypertrophic responses in primary NRCMs. Cells were transfected with plasmids for wild-type NOR1 (NOR1), a mutant of NOR1 (C293Y) and vector. (A) Expression of NOR1 protein was measured by Western blot. (B) The mRNA levels of ANF and BNP were detected by qRT-PCR. (C) Cell surface area was measured by rhodamine-phalloidin staining. Data were presented as means ± SE. *P < 0.05 versus vector group, n = 5 independent experiments.

Article Snippet: For immunoprecipitation, rabbit anti-NOR1 polyclonal antibody (diluted 1:10) and mouse anti-GFP monoclonal antibody (diluted 1:10) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

Techniques: Over Expression, Transfection, Mutagenesis, Plasmid Preparation, Expressing, Western Blot, Quantitative RT-PCR, Staining

Journal: British journal of pharmacology

Article Title: The orphan receptor NOR1 participates in isoprenaline-induced cardiac hypertrophy by regulating PARP-1.

doi: 10.1111/bph.13091

Figure Lengend Snippet: Figure 4 NOR1 physically interacted with PARP-1 and the interaction was significantly increased by isoprenaline (ISO) treatment. (A) Primary NRCMs were incubated with 1 μM isoprenaline for 24 h. Co-IP assay was performed to show the interaction between NOR1 and PARP-1 after immunoprecipitation of cardiomyocyte nuclear extracts with anti-NOR1 antibody. (B) SD rats were treated with 1.5 mg·kg−1·d−1

Article Snippet: For immunoprecipitation, rabbit anti-NOR1 polyclonal antibody (diluted 1:10) and mouse anti-GFP monoclonal antibody (diluted 1:10) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

Techniques: Incubation, Co-Immunoprecipitation Assay, Immunoprecipitation

Journal: British journal of pharmacology

Article Title: The orphan receptor NOR1 participates in isoprenaline-induced cardiac hypertrophy by regulating PARP-1.

doi: 10.1111/bph.13091

Figure Lengend Snippet: Figure 5 The activity of PARP-1 was regulated by NOR1 in cardiomyocyte hypertrophy. (A and B) PARP-1 expression and activity were measured in primary NRCMs treated with 1 μM isoprenaline (ISO) for the indicated time. (C and D) SD rats were treated with 1.5 mg·kg−1·d−1 isoprenaline or NS for 7 d. PARP-1 expression and activity were measured. (E and F) Cardiomyocytes were transfected with NOR1 siRNA (siNOR1) or negative control (NC), and further treated with 1 μM isoprenaline for 24 h. PARP-1 expression and activity were measured. (G and H) PARP-1 expression and activity were measured in cardiomyocytes overexpressing NOR1 or its mutant C293Y. Data are presented as means ± SE. *P < 0.05, versus control, normal saline (NS), NC, or vector, #P < 0.05 versus NC treated with isoprenaline, n = 5 independent experiments.

Article Snippet: For immunoprecipitation, rabbit anti-NOR1 polyclonal antibody (diluted 1:10) and mouse anti-GFP monoclonal antibody (diluted 1:10) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

Techniques: Activity Assay, Expressing, Transfection, Negative Control, Mutagenesis, Control, Saline, Plasmid Preparation

Journal: British journal of pharmacology

Article Title: The orphan receptor NOR1 participates in isoprenaline-induced cardiac hypertrophy by regulating PARP-1.

doi: 10.1111/bph.13091

Figure Lengend Snippet: Figure 7 PARP-1 inhibition attenuated hypertrophic responses induced by NOR1 overexpression. (A and B) Cardiomyocytes were transfected with PARP-1 siRNA (siPARP-1) and negative control (NC), or incubated with 3AB (500 μM) before transfection with NOR1 for 36 h. Hypertrophic responses were measured by the mRNA levels of hypertrophic biomarkers and cell surface area. Data were presented as means ± SE. *P < 0.05 versus vector, #P < 0.05 versus NOR1, n = 5 independent experiments.

Article Snippet: For immunoprecipitation, rabbit anti-NOR1 polyclonal antibody (diluted 1:10) and mouse anti-GFP monoclonal antibody (diluted 1:10) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

Techniques: Inhibition, Over Expression, Transfection, Negative Control, Incubation, Plasmid Preparation