antibody clone agr 021  (Alomone Labs)

Journal: Theranostics

Article Title: Activation of GLP-1 Receptor Enhances Neuronal Base Excision Repair via PI3K-AKT-Induced Expression of Apurinic/Apyrimidinic Endonuclease 1

doi: 10.7150/thno.15993

Figure Lengend Snippet: Activation of cortical neuron expressed GLP-1 receptor (GLP-1R) protected cortical neurons from oxidative insults. (A) mRNA and (B) protein expressions of GLP-1R were detected in the 8-day in vitro primary cortical neurons. (C) Immunofluorescence staining of GLP-1R (green), TUJ-1 (red), and DAPI (blue) in the 8-day in vitro primary cortical neurons. Merged bright field (100X) and immunostained (100X) images indicated that GLP-1R is mostly distributed in neuronal bodies but also exist in both axons and dendrites. (D) GLP-1or (E) EX-4 pretreated cortical neurons were continually treated with 100 nM GLP-1 or EX-4 after an oxidative insult and neuronal viability was measured at 0, 6, 12, 24 hr time points. Neuronal viability of ligand administrated neurons (GLP-1:~82%, EX-4:~67%) was approximately 20% higher than the menadione-treated group (GLP-1~65%, EX-4:~48%) 24-hour after menadione-induced oxidative insults. (M±SE; * p<0.05, ** p< 0.01, *** p< 0.001; compared to the 24-hr value of the menadione alone group, n=6).

Article Snippet: The dilution factors for the primary antibodies were as following, APE1 (1:1,000; Santa Cruz), phosphor-AKT (1:1,000; Cell Signaling), phosphor-CREB (1:1,000; Cell Signaling), Ligase III (1:1,000; Santa Cruz), FEN1(1:1,000; Cell Signaling), Pol β (1:200; GeneTex), NEIL1 (1:1,000; Santa Cruz), OGG1 (1:1,000; Novus), UDG (1:1,000; Novus), lamin A/C (1:1,000; Cell Signaling), GLP-1R (1:1,000; Abcam/Alomone Lab) and actin (1:5,000; Sigma-Aldrich).

Techniques: Activation Assay, In Vitro, Immunofluorescence, Staining

Journal: Theranostics

Article Title: Activation of GLP-1 Receptor Enhances Neuronal Base Excision Repair via PI3K-AKT-Induced Expression of Apurinic/Apyrimidinic Endonuclease 1

doi: 10.7150/thno.15993

Figure Lengend Snippet: The downstream PI3K-AKT signaling axis of GLP-1R is the major pathway that regulates expression of APE1. (A) Western blot analyses of pAKT, pERK1/2, pCREB, APE1, and actin levels in neuronal cultures treated with 100 nM of GLP-1(100nM), GLP-1 plus MEK inhibitors U0126, or GLP-1 plus PI3K inhibitor LY294002. (B) LY294002 specifically inhibited GLP-1-induced AKT phosphorylation. (C) U0126 specifically inhibited GLP-1-induced pERK1/2 phosphorylation. (D) pCREB levels and (E) APE1 were specifically inhibited by LY294002 but not U0126. The results suggested that PI3K-AKT-CREB is the predominant downstream signaling pathway of GLP-1R elevating APE1 expression. (M±SE; *p< 0.05; ** p< 0.01; ***p<0.001; compared to the value of control group, n=4).

Article Snippet: The dilution factors for the primary antibodies were as following, APE1 (1:1,000; Santa Cruz), phosphor-AKT (1:1,000; Cell Signaling), phosphor-CREB (1:1,000; Cell Signaling), Ligase III (1:1,000; Santa Cruz), FEN1(1:1,000; Cell Signaling), Pol β (1:200; GeneTex), NEIL1 (1:1,000; Santa Cruz), OGG1 (1:1,000; Novus), UDG (1:1,000; Novus), lamin A/C (1:1,000; Cell Signaling), GLP-1R (1:1,000; Abcam/Alomone Lab) and actin (1:5,000; Sigma-Aldrich).

Techniques: Expressing, Western Blot

Journal: Theranostics

Article Title: Activation of GLP-1 Receptor Enhances Neuronal Base Excision Repair via PI3K-AKT-Induced Expression of Apurinic/Apyrimidinic Endonuclease 1

doi: 10.7150/thno.15993

Figure Lengend Snippet: This schematic diagram illustrates the mechanisms by which activated GLP-1R elevate repair of oxidative DNA damage and increase neuronal survival. The expression GLP-1 and its receptors occurs in the central nervous system and is involved in neuronal protection. Two major signaling pathways are triggered, including PKA-Erk (dotted-line arrow) and PI3K-AKT (filled-line arrow), when GLP-1R is activated. Results of our study suggested that PI3K-AKT is the key downstream signaling pathway regulating APE1 expression, elevation DNA repair efficiency, and increasing neuronal survival after oxidative insults. (AC: adenylyl cyclase; cAMP: cyclic adenosine monophosphate; PKA: protein kinase A; MEK: mitogen-activated protein kinase kinase; ERK: mitogen-activated protein kinase; PI3K: phosphatidylinositide 3-kinase; AKT: protein kinase B; CREB: cAMP response element binding protein).

Article Snippet: The dilution factors for the primary antibodies were as following, APE1 (1:1,000; Santa Cruz), phosphor-AKT (1:1,000; Cell Signaling), phosphor-CREB (1:1,000; Cell Signaling), Ligase III (1:1,000; Santa Cruz), FEN1(1:1,000; Cell Signaling), Pol β (1:200; GeneTex), NEIL1 (1:1,000; Santa Cruz), OGG1 (1:1,000; Novus), UDG (1:1,000; Novus), lamin A/C (1:1,000; Cell Signaling), GLP-1R (1:1,000; Abcam/Alomone Lab) and actin (1:5,000; Sigma-Aldrich).

Techniques: Expressing, Binding Assay