Journal: Scientific Reports

Article Title: Activation of PPAR-α attenuates myocardial ischemia/reperfusion injury by inhibiting ferroptosis and mitochondrial injury via upregulating 14-3-3η

doi: 10.1038/s41598-024-64638-9

Figure Lengend Snippet: Ppar-α inhibits ferroptosis of A/R-induced H9c2 cells by downregulating 14-3-3η. The concentrations of ( A ) MDA, ( B ) total iron, and ( C , D ) GSH and GSSG were determined by corresponding quantitative kits in A/R-induced cells after GW7647, GW6471, or Fer-1 pretreatment. The levels of ( E , F ) ROS and ( G , I ) ferrous iron were measured by corresponding quantitative kits in A/R-induced cells after GW7647, GW6471, or Fer-1 induced (magnification, ×200). ( H , J ) The levels of Lipid ROS was assessed by C11-BODIPY staining. ( K , L ) the expression of 14-3-3η, Ppar-α, and ferroptosis-related proteins were detected by Western blot in A/R-induced cells after GW7647, GW6471, or Fer-1 pretreatment. Data are expressed as the mean ± SD (n = 3). ** P < 0.05.

Article Snippet: Mitochondrial permeability transition pore (mPTP), cell apoptosis, and the levels of ROS and lipid ROS were determined by the mPTP detection kit (BestBio, Shanghai, China), Annexin V-FITC apoptosis detection Kit (BestBio, Shanghai, China), ROS assay kit (Beyotime, Shanghai, China), and lipid ROS assay kit (GLPBIO, Montclair, CA, USA) according to the protocol provided with the corresponding assay kit.

Techniques: Staining, Expressing, Western Blot

Journal: Scientific Reports

Article Title: Activation of PPAR-α attenuates myocardial ischemia/reperfusion injury by inhibiting ferroptosis and mitochondrial injury via upregulating 14-3-3η

doi: 10.1038/s41598-024-64638-9

Figure Lengend Snippet: Ppar-α inhibits ferroptosis of A/R-induced H9c2 cells by mediating 14-3-3η. ( A ) CCK-8 detected the cell viability in A/R-induced H9c2 cells after GW7647, pAd/14-3-3η-shRNA, and pAd/NC-shRNA pretreatment. The concentrations of ( B ) LDH activities, ( C ) MDA, ( D ) total iron, and ( E ) GSH, GSSG, and GSH/GSSG were determined by corresponding quantitative kits in A/R-induced cells after GW7647, pAd/14-3-3η -shRNA, and pAd/NC-shRNA induced. ( F , H ) the concentrations of ferrous iron were measured by corresponding quantitative kits in A/R-induced cells after GW7647, pAd/14-3-3η-shRNA, and pAd/NC-shRNA pretreatment (magnification, ×200). ( G , I ) The levels of Lipid ROS was assessed by C11-BODIPY staining. ( J , K ) the expression of 14-3-3η, Ppar-α, and ferroptosis-related proteins were detected by Western blot in A/R-induced cells after GW7647, pAd/14-3-3η-shRNA, and pAd/NC-shRNA induced. Data are expressed as the mean ± SD (n = 3). ** P < 0.05.

Article Snippet: Mitochondrial permeability transition pore (mPTP), cell apoptosis, and the levels of ROS and lipid ROS were determined by the mPTP detection kit (BestBio, Shanghai, China), Annexin V-FITC apoptosis detection Kit (BestBio, Shanghai, China), ROS assay kit (Beyotime, Shanghai, China), and lipid ROS assay kit (GLPBIO, Montclair, CA, USA) according to the protocol provided with the corresponding assay kit.

Techniques: CCK-8 Assay, shRNA, Staining, Expressing, Western Blot