Journal: Experimental Animals

Article Title: Acyl-CoA thioesterase 1 (ACOT1) overexpression alleviates heart failure by inhibiting oxidative stress and cardiomyocyte apoptosis through the Kelch-like ECH-associated protein1-NF-E2-related factor2 (KEAP1-NRF2) pathway

doi: 10.1538/expanim.24-0129

Figure Lengend Snippet: (A) The levels of MDA and GSH and the activity of SOD in myocardial tissues were analyzed by ELISA. (B) The protein levels and the quantification results of cleaved PARP, cleaved CASPASE-3 and cleaved CASPASE-9 in myocardial tissues. (C) ROS in myocardial tissues were detected by DHE staining (n=6). The fluorescence intensity was calculated to evaluate ROS levels. bar=50 µ m. # P <0.05, ## P <0.01, ### P <0.001. n=6. EP: empty plasmid; ACOT1 oe : ACOT1 overexpression.

Article Snippet: The following antibodies were used: cleaved CASPASE-3 (Affinity, Cincinnati, OH, USA, #AF7022) diluted in 1:1,000; cleaved CASPASE-9 (Affinity, #AF5240) diluted in 1:1,000; cleaved PARP (ABclonal, Wuhan, China, #A22535) diluted in 1:1,000; ACOT1 (Novus Biologicals, Littleton, CO, USA, #NBP3-10843) diluted in 1:500; KEAP1 (Affinity, #AF5266) diluted in 1:1,000; NRF2 (Affinity, #AF0639) diluted in 1:1,000; Histone H3 (Gene Tex, Alton Parkway Irvine, CA, USA, #GTX122148) diluted in 1:5,000; GAPDH (Proteintech, Wuhan, China, #60004-1-Ig) diluted in 1:10,000; Goat Anti-Rabbit IgG/HRP (Solarbio, #SE134) diluted in 1:3,000; Goat Anti-Mouse IgG/HRP (Solarbio, #SE131) diluted in 1:3,000.

Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay, Staining, Fluorescence, Plasmid Preparation, Over Expression

Journal: Experimental Animals

Article Title: Acyl-CoA thioesterase 1 (ACOT1) overexpression alleviates heart failure by inhibiting oxidative stress and cardiomyocyte apoptosis through the Kelch-like ECH-associated protein1-NF-E2-related factor2 (KEAP1-NRF2) pathway

doi: 10.1538/expanim.24-0129

Figure Lengend Snippet: (A) Protein level and the quantification of ACOT1 in HL-1 cells with ACOT1 overexpression. (B) cell viability of HL-1 cells with hypoxia treatment. (C) The levels of MDA and GSH and the activity of SOD in cells. (D) The protein levels of cleaved PARP, cleaved CASPASE-3 and cleaved CASPASE-9 in cells and the quantification results. (E) TUNEL staining of HL-1 cells and the positive cell quantification, bar=100 µ m. (F) ROS levels in HL-1 cells were detected by flow cytometry, MFI: mean fluorescence intensity. ## P <0.01, ### P <0.001. n=3. EP: empty plasmid; ACOT1 oe : ACOT1 overexpression.

Article Snippet: The following antibodies were used: cleaved CASPASE-3 (Affinity, Cincinnati, OH, USA, #AF7022) diluted in 1:1,000; cleaved CASPASE-9 (Affinity, #AF5240) diluted in 1:1,000; cleaved PARP (ABclonal, Wuhan, China, #A22535) diluted in 1:1,000; ACOT1 (Novus Biologicals, Littleton, CO, USA, #NBP3-10843) diluted in 1:500; KEAP1 (Affinity, #AF5266) diluted in 1:1,000; NRF2 (Affinity, #AF0639) diluted in 1:1,000; Histone H3 (Gene Tex, Alton Parkway Irvine, CA, USA, #GTX122148) diluted in 1:5,000; GAPDH (Proteintech, Wuhan, China, #60004-1-Ig) diluted in 1:10,000; Goat Anti-Rabbit IgG/HRP (Solarbio, #SE134) diluted in 1:3,000; Goat Anti-Mouse IgG/HRP (Solarbio, #SE131) diluted in 1:3,000.

Techniques: Over Expression, Activity Assay, TUNEL Assay, Staining, Flow Cytometry, Fluorescence, Plasmid Preparation

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Deferasirox Targeting Ferroptosis Synergistically Ameliorates Myocardial Ischemia Reperfusion Injury in Conjunction With Cyclosporine A

doi: 10.1161/JAHA.123.031219

Figure Lengend Snippet: A, Experimental protocol for inducing ischemia reperfusion (I/R) in mice. B, Measurement of total myocardial iron levels 24 hours after DFX administration (200 mg/kg, per os [p.o.]; n=15, each group). C, Measurement of iron levels in the ER fraction of I/R‐injured myocardium 24 hours after I/R (n=4, each group). D, Measurement of iron levels in the ER fraction of I/R‐injured myocardium treated with vehicle or DFX (200 mg/kg, per os [p.o.]; n=8 in vehicle group and 7 in DFX group). Data are presented as the mean±SEM. Statistical significance was determined using unpaired t ‐test. * P <0.05, ** P <0.01. 4‐HNE indicates 4‐hydroxynonenal; CCM, cleaved caspase substrate motif; HO‐1, heme oxygenase‐1; Hx, hypoxia; LAD, left anterior descending artery; Nx, normoxia; TUNEL, terminal deoxynucleotidyl transferase‐mediated dUTP nick end labeling; and Veh, vehicle.

Article Snippet: Western blotting was performed as described previously , with the following primary antibodies against: 4‐hydroxy‐2‐nonenal (4‐HNE; MHN‐020P, JaICA; Shizuoka), acrolein (MAR‐020n, JaICA), cyclooxygenase‐2 (COX2; #12282, Cell Signaling Technology [CST]), heme oxygenase‐1 (HO‐1; #43966, CST), cleaved caspase substrate motif (#8698, CST), BAX (#14796, CST), and BCL2 (ab182858, Abcam); the antibody against GAPDH (internal standard; Santa Cruz Biotechnology, Inc., sc‐32 233) was used as an internal control to standardize the signal intensities.

Techniques: TUNEL Assay, End Labeling