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

Journal: International Journal of Molecular Sciences

Article Title: Polyoxometalate K 6 [P 2 Mo 18 O 62 ] Inactivates Escherichia coli O157:H7 by Inducing recA Expression and Apoptosis-like Bacterial Death

doi: 10.3390/ijms241411469

Figure Lengend Snippet: Polyoxometalate K 6 [P 2 Mo 18 O 62 ] treatment induced characteristic apoptosis-like bacterial death events. ( A ) Quantitation of TUNEL staining by flow cytometry. ( B ) Quantitation of annexin V-FITC/propyl iodide staining by flow cytometry.

Article Snippet: The bacteria were then washed twice with PBS and labelled using a One Step TUNEL Apoptosis Assay Kit (containing a mixture of terminal deoxynucleotidyl transferase and FITC-conjugated dUTP) (Beyotime, Shanghai, China).

Techniques: Quantitation Assay, TUNEL Assay, Staining, Flow Cytometry

Journal: Frontiers in Cell and Developmental Biology

Article Title: BRG1 Links TLR4 Trans-Activation to LPS-Induced SREBP1a Expression and Liver Injury

doi: 10.3389/fcell.2021.617073

Figure Lengend Snippet: BRG1 deficiency in hepatocyte alleviates LPS-induced liver injury. WT and BRG1 LKO mice were injected peritoneally with LPS (15 mg/kg) and sacrificed 24 h after the injection. (A) Plasma ALT levels. N = 5 mice for the saline groups and N = 8 mice for the LPS groups. (B) Plasma AST levels. N = 5 mice for the saline groups and N = 8 mice for the LPS groups. (C) Paraffin sections were stained with F4/80. N = 6 mice for each group. (D) Paraffin sections were stained with TUNEL. N = 6 mice for each group. (E) Gene expression levels were examined by qPCR. N = 6 mice for each group.

Article Snippet: Briefly, paraffin sections were incubated in the TUNEL reaction mixture (R&D Systems, 4810-30-K) at 37°C for 60 min. After several rinses with PBS, the sections were incubated with an anti-HRP antibody at room temperature for 30min.

Techniques: Injection, Clinical Proteomics, Saline, Staining, TUNEL Assay, Gene Expression

Journal: Frontiers in Cell and Developmental Biology

Article Title: BRG1 Links TLR4 Trans-Activation to LPS-Induced SREBP1a Expression and Liver Injury

doi: 10.3389/fcell.2021.617073

Figure Lengend Snippet: TLR4 over-expression or SREBP1a over-expression restores liver injury in BRG1 deficient mice. (A–F) WT and BRG1 LKO mice were injected via tail veinadenovirus carrying either TLR4 (Ad-GFP-TLR4), SREBP1a (Ad-GFP-SREBP1a), or GFP (Ad-GFP). 2 week later, the mice were injected with injected peritoneally with LPS (15 mg/kg) and sacrificed 24 h after the injection. Gene expression levels were examined by Western (A) . Plasma ALT levels (B) . Plasma AST levels (C) . Paraffin sections were stained with F4/80 (D) . Paraffin sections were stained with TUNEL (E) . Gene expression levels were examined by qPCR (F) . N = 6 mice for all groups. (G) Primary hepatocytes were isolated from WT or BRG1 LKO mice and transduced with indicated andenoviral particles. The cells were with LPS (1 μg/ml) for 12 h and gene expression was examined by qPCR.

Article Snippet: Briefly, paraffin sections were incubated in the TUNEL reaction mixture (R&D Systems, 4810-30-K) at 37°C for 60 min. After several rinses with PBS, the sections were incubated with an anti-HRP antibody at room temperature for 30min.

Techniques: Over Expression, Injection, Gene Expression, Western Blot, Clinical Proteomics, Staining, TUNEL Assay, Isolation, Transduction