rabbit polyclonal anti pp2a c subunit  (Cell Signaling Technology Inc)

Journal: Cell metabolism

Article Title: Fructose-1,6-bisphosphatase is a nonenzymatic safety valve that curtails AKT activation to prevent insulin hyperresponsiveness

doi: 10.1016/j.cmet.2023.03.021

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit Polyclonal anti-PP2A-C , Cell Signaling , Cat# 2038; RRID:AB_2169495.

Techniques: Immunoprecipitation, Transfection, Extraction, Enzyme-linked Immunosorbent Assay, ATP Assay, Colorimetric Assay, Activity Assay, Glucose Assay, In Situ, cDNA Synthesis, Recombinant, Sequencing, Construct, Software

Journal: Molecular Neurobiology

Article Title: Cancerous Inhibitor of Protein Phosphatase 2A (CIP2A): Could It Be a Promising Biomarker and Therapeutic Target in Parkinson’s Disease?

doi: 10.1007/s12035-021-02670-w

Figure Lengend Snippet: Assessment of CIP2A expression levels in rotenone treated PD cell models. a, b The IF staining of p-α-syn (green) increased following incubation of the SH-SY5Y cells with 0.25 uM and 0.5 uM rotenone for 24 h. c–e IF staining showed increased CIP2A (green) expression in PD group and colocalized of α-syn (red) with CIP2A. f Western blotting results of CIP2A , PP2A, and p-α-syn revealed the ascending CIP2A and p-α-syn after rotenone treatment; β-actin served as the loading control. g–i Quantification of CIP2A/GAPDH, PP2A/GAPDH, or p-α-syn/GAPDH ratio, the bars represent the mean ± SEM. Histograms summarize the mean IF signal intensity ± SEM as measured in gray values. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001

Article Snippet: Rotenone and MPTP were purchased from Sigma-Aldrich (R8875, M0896, St. Louis, MO, USA); rabbit polyclonal α-syn phospho-S129 antibody, sheep polyclonal α-syn antibody, and sheep polyclonal tyrosine hydroxylase (TH) antibody were obtained from Abcam (ab51253, ab6162, ab113, UK); rabbit polyclonal PP2A and CIP2A antibodies were from CST (2038 T, 14805S, MA, USA); rabbit polyclonal CIP2A antibody was obtained from ABclonal (A12267, Wuhan, China), while rabbit polyclonal α-syn antibody was purchased from Genetex (GTX112799, San Antonio, TX, USA). siRNA for CIP2A was designed by RiboBio Co., Ltd., Guangzhou, China, as follows: siCIP2A1 CTGTGGTTGTGTTTGCACT; siCIP2A2 GCTCTACTGCGCTGGTTAA; siCIP2A3 CACGGACACTTGCTAGTAT.

Techniques: Expressing, Staining, Incubation, Western Blot, Control

Journal: Molecular Neurobiology

Article Title: Cancerous Inhibitor of Protein Phosphatase 2A (CIP2A): Could It Be a Promising Biomarker and Therapeutic Target in Parkinson’s Disease?

doi: 10.1007/s12035-021-02670-w

Figure Lengend Snippet: Assessment of the effect of CIP2A knockdown on rotenone induced PD pathology. a , c Western blotting assays were used to evaluate the interference effects of the three siRNAs on the CIP2A expression. b , d IF staining of CIP2A (green) confirmed the effect of CIP2A knockdown by siCIP2A1. e , f , g , h Representative IF staining of the p-α-syn and α-syn in the control, siRNA-treated, rotenone-treated, or siRNA + rotenone-treated group. i The PP2A activity was decreased after rotenone exposure but increased after siCIP2A treatment. k, j The α-syn expression decreased after treatment with the three pairs of CIP2A siRNAs. l , n CIP2A interacts directly with α-syn. Co-IP and western blot analysis of the CIP2A and α-syn from the SH-SY5Y cells; normal rabbit IgG served as the controls. m Confocal fluorescence microscopy showing colocalization of CIP2A and α-syn. Values are given as means ± SEM. Statistical significance was determined by one-way ANOVA. Histograms summarize the mean IF signal intensity detected ± SEM as measured in gray values. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001

Article Snippet: Rotenone and MPTP were purchased from Sigma-Aldrich (R8875, M0896, St. Louis, MO, USA); rabbit polyclonal α-syn phospho-S129 antibody, sheep polyclonal α-syn antibody, and sheep polyclonal tyrosine hydroxylase (TH) antibody were obtained from Abcam (ab51253, ab6162, ab113, UK); rabbit polyclonal PP2A and CIP2A antibodies were from CST (2038 T, 14805S, MA, USA); rabbit polyclonal CIP2A antibody was obtained from ABclonal (A12267, Wuhan, China), while rabbit polyclonal α-syn antibody was purchased from Genetex (GTX112799, San Antonio, TX, USA). siRNA for CIP2A was designed by RiboBio Co., Ltd., Guangzhou, China, as follows: siCIP2A1 CTGTGGTTGTGTTTGCACT; siCIP2A2 GCTCTACTGCGCTGGTTAA; siCIP2A3 CACGGACACTTGCTAGTAT.

Techniques: Knockdown, Western Blot, Expressing, Staining, Control, Activity Assay, Co-Immunoprecipitation Assay, Fluorescence, Microscopy

Journal: Molecular Neurobiology

Article Title: Cancerous Inhibitor of Protein Phosphatase 2A (CIP2A): Could It Be a Promising Biomarker and Therapeutic Target in Parkinson’s Disease?

doi: 10.1007/s12035-021-02670-w

Figure Lengend Snippet: Evaluation of CIP2A expression in MPTP induced subacute mouse models. a, l Representative IF images and the quantification of the number of TH + neurons in the SN at 14 days after treatment with MPTP, n = 4 to 6 brains per group. b Behavioral test of the mice showed decreased time in rotarod test and extended time in pole test after MPTP treatment. Data were expressed as means ± SEM, n = 8 to 9 mice per group. c Compared with the control group, there was suppression of the PP2A activity of the brain tissue of the MPTP-treated mice, n = 6 brains per group. d , o Immunoblot results demonstrating the decreased TH expression in both the SN and striatum of the MPTP-treated mice; β-actin served as the loading control. e , p Immunoblot results demonstrating CIP2A expression in the SN of the mice. f Immunoblotting of α-syn . g , h , m , n IF staining of CIP2A (green) and TH (red) as well as IHC assay showed increased CIP2A expression in the SN after MPTP treatment, n = 4 to 6 brains per group. i Plasma CIP2A levels were significantly decreased in the MPTP treated mice when compared to those in the controls. j Immunoblotting of p-α-syn in the SN of rotenone model. k IHC staining of TH and CIP2A in the SN of rotenone model. Histograms summarize the mean IF signal intensity detected ± SEM as measured in gray values. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001

Article Snippet: Rotenone and MPTP were purchased from Sigma-Aldrich (R8875, M0896, St. Louis, MO, USA); rabbit polyclonal α-syn phospho-S129 antibody, sheep polyclonal α-syn antibody, and sheep polyclonal tyrosine hydroxylase (TH) antibody were obtained from Abcam (ab51253, ab6162, ab113, UK); rabbit polyclonal PP2A and CIP2A antibodies were from CST (2038 T, 14805S, MA, USA); rabbit polyclonal CIP2A antibody was obtained from ABclonal (A12267, Wuhan, China), while rabbit polyclonal α-syn antibody was purchased from Genetex (GTX112799, San Antonio, TX, USA). siRNA for CIP2A was designed by RiboBio Co., Ltd., Guangzhou, China, as follows: siCIP2A1 CTGTGGTTGTGTTTGCACT; siCIP2A2 GCTCTACTGCGCTGGTTAA; siCIP2A3 CACGGACACTTGCTAGTAT.

Techniques: Expressing, Control, Activity Assay, Western Blot, Staining, Clinical Proteomics, Immunohistochemistry