Journal: Cell Death & Disease
Article Title: SIRT1 deficiency promotes age-related heart failure through enhancing ferroptosis via GATA4-HADHA-GPX4 axis
doi: 10.1038/s41419-026-08634-z
Figure Lengend Snippet: A Neonatal rat cardiomyocytes were transfected with negative control siRNA (si-NC) or siRNA targeting SIRT1 (si-SIRT1). Representative immunoblots and quantitative analysis of SIRT1, HADHA, transferrin (TF), ferritin heavy chain 1 (FTH1), and glutathione peroxidase 4 (GPX4) protein levels are shown ( n = 5 per group). B In silico molecular docking analysis illustrating the predicted interaction between SIRT1 and HADHA. C Relative mRNA expression of Hadha in cardiomyocytes following SIRT1 silencing, as determined by quantitative real-time PCR ( n = 6 per group). D Bioinformatic enrichment analysis identifying GATA4 as a putative transcription factor associated with HADHA regulation, based on the ChEA Transcription Factor Targets and Binding Site Profiles datasets. E Co-immunoprecipitation analysis demonstrating the interaction between SIRT1 and GATA4 in cardiomyocytes. Cell lysates were immunoprecipitated with anti-SIRT1 antibody, followed by immunoblotting with antibodies against SIRT1 and GATA4. Representative immunoblots are shown. F Schematic illustration of the HADHA promoter region showing the predicted GATA4-binding motif (−327 to −337 bp) and the serial 5′-truncated promoter constructs generated for luciferase reporter assays. ( G ) Dual-luciferase reporter assays assessing the transcriptional activity of individual serially truncated HADHA promoter constructs following GATA4 overexpression or control treatment ( n = 6 per group). H Experimental design for resveratrol intervention in aging rats. Eighteen-month-old rats were randomly assigned to receive vehicle (Aging Control) or resveratrol (Aging RES) by daily gavage for 4 months and were analyzed at 22 months of age. I Representative immunoblots of SIRT1, HADHA, and GATA4 protein expression in cardiac tissues from Aging Control and Aging RES rats. J Representative immunoblots of TF, FTH1, and GPX4 protein expression in cardiac tissues from Aging Control and Aging RES rats. K Quantitative analysis of SIRT1, HADHA, GATA4, TF, FTH1, and GPX4 protein levels in cardiac tissues from Aging Control and Aging RES rats ( n = 6 per group). L Representative M-mode echocardiographic images from Aging Control and Aging RES rats. M Left ventricular ejection fraction (EF) in Aging Control and Aging RES rats ( n = 6 per group). N Left ventricular fractional shortening (FS) in Aging Control and Aging RES rats ( n = 6 per group). O Serum NT-proBNP levels in Aging Control and Aging RES rats ( n = 6 per group). P Representative hematoxylin and eosin (H&E)–stained sections of left ventricular tissue. Scale bars, 50 μm. Q Representative Masson trichrome–stained sections of left ventricular tissue. Scale bars, 50 μm. R Quantification of left ventricular collagen volume fraction ( n = 6 per group). The data are given as mean ± SEM and compared by Student’s t test.
Article Snippet: The membranes were then incubated with primary antibodies against TF (1:500, Proteintech, cat#17435-1-AP, RRID: AB_2035023), FTH1 (1:500, Bioss, cat#bs-5907R, RRID: AB_11050926), GPX4 (1:500, Abcam, cat#ab125066, RRID: AB_10973901), HADHA (1:2000, Proteintech, cat#10758-1-AP, RRID: AB_2115593), and SIRT1 (IP, 1:500, Proteintech, cat#60303-1-Ig, RRID: AB_2881417; IB, 1:500, Abcam, cat#ab189494, RRID: AB_2864311), GATA4 (1:500, Proteintech, cat#19530-1-AP, RRID: AB_10642003), and GAPDH (1:10000, Abcam, cat#ab128915, RRID: AB_2747414) at 4°C overnight.
Techniques: Transfection, Negative Control, Western Blot, In Silico, Expressing, Real-time Polymerase Chain Reaction, Binding Assay, Immunoprecipitation, Construct, Generated, Luciferase, Activity Assay, Over Expression, Control, Staining
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