anti sglt2  (Alomone Labs)

Journal: International Journal of Molecular Sciences

Article Title: Dapagliflozin Prevents High-Glucose-Induced Cellular Senescence in Renal Tubular Epithelial Cells

doi: 10.3390/ijms232416107

Figure Lengend Snippet: High glucose increased glucose consumption and SGLT2 expression, while dapagliflozin decreased both. In high-glucose conditions, glucose consumption was increased. Dapagliflozin reduced glucose consumption under normal and high-glucose conditions ( A ). High glucose upregulated SGLT2 expression. Dapagliflozin decreased SGLT-2 expression under normal and high-glucose conditions ( B , C ). * p < 0.05 vs. control; # p < 0.05 vs. RPTECs treated with 15 ng/mL dapagliflozin; ^ p < 0.05 vs. RPTECs in high glucose; + p < 0.05 vs. RPTECs in high glucose and treated with 15 ng/mL dapagliflozin. SGLT-2, sodium–glucose cotransporter-2.

Article Snippet: The primary antibodies were specific for the following proteins: SGLT-2 (1:200, cat. No AGT-032, Alomone Labs, Jerusalem, Israel), 4-Hydroxynonenal (4-HNE, 1:500, cat. No ab46545 Abcam, Cambridge, UK), ataxia telangiectasia mutated kinase (ATM, 1:1000, cat. No 2873, Cell Signaling Technology, Danvers, MA, USA), phosphorylated at Ser1981 ATM (p-ATM, 1:1000, cat. No 5883, Cell Signaling Technology); tumor suppressor p53 (p53, 1:1000, cat. No 2524, Cell Signaling Technology), phosphorylated at Ser15 p53 (p-p53, 1:1000, cat. No 9284, Cell Signaling Technology), phosphorylated at Ser139 histone H2AX (γ-H2AX, 1:500, cat. No NB100-2280, Novus Biologicals, Abingdon, Oxon, UK), p21 Waf1/Cip1 (p21, 1:1000, cat. No 37543, Cell Signaling Technology), p16 INK4A (p16, 1:1000, cat. No 80772, Cell Signaling Technology), marker of proliferation Ki-67 (Ki-67, 1:1000, cat no NBP2-22112, Novus Biologicals), beta-galactosidase (GLB-1, 1:500, cat. No ab55176, Abcam), α-smooth muscle actin (α-SMA, 1:100, cat no. sc-130617; Santa Cruz Biotechnology, Dallas, TC, USA), and β-actin (1:2500, cat. no 4967, Cell Signaling Technology).

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: Dapagliflozin Prevents High-Glucose-Induced Cellular Senescence in Renal Tubular Epithelial Cells

doi: 10.3390/ijms232416107

Figure Lengend Snippet: Dapagliflozin prevents high-glucose-induced RPTEC senescence. High glucose increased SGLT-2 expression and glucose consumption, enhancing ROS production. The latter induced DNA damage, ATM, and p53 phosphorylation. Stabilized p53 increased the cell cycle inhibitor p21, resulting in cell cycle arrest and increasing the cellular senescence marker GLB-1. In addition, RPTECs acquired am SASP phenotype, which was detected by the production of IL-1β, IL-8, and TGF-β1. By decreasing SGLT-2 expression and glucose consumption, dapagliflozin inhibited the above pathway and prevented RPTEC senescence. In addition, dapagliflozin reduced the cell cycle inhibitor p16 independently of the glucose conditions. 4-HNE, 4-Hydroxynonenal; γ-H2AX, phosphorylated at Ser139 histone H2AX; ATM, ataxia telangiectasia mutated kinase; GLB-1, beta-galactosidase; IL-1β, interleukin-1β; IL-8, interleukin-8; Ki-67, marker of proliferation Ki-67; p16, p16 INK4A, p21, p21 Waf1/Cip1; p53, tumor suppressor p53; ROS, reactive oxygen species; RPTEC, renal proximal tubular epithelial cell; SASP, senescence-associated secretory phenotype; SGLT-2, sodium–glucose cotransporter-2; TGF-β1, transforming growth factor-β1.

Article Snippet: The primary antibodies were specific for the following proteins: SGLT-2 (1:200, cat. No AGT-032, Alomone Labs, Jerusalem, Israel), 4-Hydroxynonenal (4-HNE, 1:500, cat. No ab46545 Abcam, Cambridge, UK), ataxia telangiectasia mutated kinase (ATM, 1:1000, cat. No 2873, Cell Signaling Technology, Danvers, MA, USA), phosphorylated at Ser1981 ATM (p-ATM, 1:1000, cat. No 5883, Cell Signaling Technology); tumor suppressor p53 (p53, 1:1000, cat. No 2524, Cell Signaling Technology), phosphorylated at Ser15 p53 (p-p53, 1:1000, cat. No 9284, Cell Signaling Technology), phosphorylated at Ser139 histone H2AX (γ-H2AX, 1:500, cat. No NB100-2280, Novus Biologicals, Abingdon, Oxon, UK), p21 Waf1/Cip1 (p21, 1:1000, cat. No 37543, Cell Signaling Technology), p16 INK4A (p16, 1:1000, cat. No 80772, Cell Signaling Technology), marker of proliferation Ki-67 (Ki-67, 1:1000, cat no NBP2-22112, Novus Biologicals), beta-galactosidase (GLB-1, 1:500, cat. No ab55176, Abcam), α-smooth muscle actin (α-SMA, 1:100, cat no. sc-130617; Santa Cruz Biotechnology, Dallas, TC, USA), and β-actin (1:2500, cat. no 4967, Cell Signaling Technology).

Techniques: Expressing, Marker