Journal: Chinese Medicine

Article Title: Fat-targeted small molecule alleviates abnormal adipose tissue remodeling in obesity via SIRT3-driven mitophagy and inflammasome inhibition

doi: 10.1186/s13020-025-01253-4

Figure Lengend Snippet: ALM blunted mitochondrial damage through SIRT3-mediated SOD2 deacetylation and mitophagy activation. A The mitochondrial reactive oxygen species (mtROS) level was determined by MitoSOX in LPS plus ATP-challenged RAW 264.7 cells treated with or without ALM and 3-TYP ( n = 5). B Mitochondrial membrane potential was evaluated by JC-1 staining ( n = 6). C Co-IP assay indicates the level of acetylated SOD2 in LPS plus ATP-challenged RAW 264.7 cells treated with or without ALM and 3-TYP ( n = 5). D RAW264.7 cells were transiently infected with the mCherry-LC3 plasmid for 24 h, and immunofluorescence staining of Mito-tracker was performed ( n = 5). E Western blot analysis of mitophagy components PINK1, Parkin, and FUNDC1 in LPS plus ATP-stimulated macrophages with the indicated concentration of ALM and 3-TYP ( n = 5). β-actin was used as an internal loading control. F The immunofluorescence staining of FUNDC1 and mCherry-LC3 was performed ( n = 5). Data are expressed as means ± SEM. G Western blot analysis of NLRP3 and Caspase1 p20 levels in LPS plus ATP-stimulated macrophages with the indicated concentration of ALM and Si-FUNDC1 ( n = 5). β-actin was used as an internal loading control ( n = 5). H–J The levels of TNF-α, MCP-1, and IL-6 in the culture medium were determined by ELISA kits ( n = 5). Data are expressed as means ± SEM. # P < 0.05 and ### P < 0.001, LPS + ATP vs. CTRL; * P < 0.05, ** P < 0.01 and *** P < 0.001, LPS + ATP + ALM vs. LPS + ATP; & P < 0.05 and && P < 0.01, LPS + ATP + ALM vs. LPS + ATP + ALM + 3-TYP or LPS + ATP + ALM + Si-FDUNC1

Article Snippet: FUNDC1 siRNA , Santa Cruz , SC-145273.

Techniques: Activation Assay, Membrane, Staining, Co-Immunoprecipitation Assay, Infection, Plasmid Preparation, Immunofluorescence, Western Blot, Concentration Assay, Control, Enzyme-linked Immunosorbent Assay

Journal: Investigative Ophthalmology & Visual Science

Article Title: ER Stress and Mitochondrial Perturbations Regulate Cell Death in Retinal Detachment: Exploring the Role of HIF1α

doi: 10.1167/iovs.65.11.39

Figure Lengend Snippet: Hypoxia and retinal detachment activate mitophagy markers in retinal cells. ( A ) Representative immunoblots of PINK1, Parkin and FUNDC1 proteins from 661W cells subjected to 8 hours, 18 hours, and 24 hours of hypoxia. Blots are representative of three independent experiments in each condition; α-tubulin and GAPDH were used as loading controls. ( B ) Representative immunocytochemistry images of LC3B ( green ) and FUNDC1 ( red ) stained 661W cells subjected to 24 hours of hypoxia compared to normoxic control. Scale bar : 64 µm. ( C ) Representative immunoblots of Parkin and FUNDC1 proteins from retinas of C57BL/6J mice 1 day and 3 days following retinal detachment. Blots are representative of three independent animals in each condition; α-tubulin was used as the loading control. ( D ) Relative mRNA expression of Pink1 obtained at 1 dprd and 3 dprd compared to the contralateral eye (fellow). Pum-1 was used as a housekeeping control gene for normalization. ( E ) Relative mRNA expression of Prkn obtained at 1 dprd and 3 dprd compared to the contralateral eye (fellow). Pum-1 was used as a housekeeping control gene for normalization. Bar graphs represent mean ± SEM. Statistical analysis was performed using one-way ANOVA with repeated measures followed by Tukey's test. * P < 0.05; ** P < 0.01.

Article Snippet: After fixation, the cells were permeabilized with 0.1% Triton X-100, blocked using 2% BSA, and subsequently subjected to an overnight incubation with the antibodies against FUNDC1 (1:250; Novus Biologicals) or anti-mouse LC3B (#83506, 1:250; Cell Signaling Technology).

Techniques: Western Blot, Immunocytochemistry, Staining, Control, Expressing

Journal: Investigative Ophthalmology & Visual Science

Article Title: ER Stress and Mitochondrial Perturbations Regulate Cell Death in Retinal Detachment: Exploring the Role of HIF1α

doi: 10.1167/iovs.65.11.39

Figure Lengend Snippet: HIF1α is associated with mitophagy induction after RD in retinal cells. ( A ) Representative immunoblots of HIF1α protein from 661W cells subjected to 8 hours, 18 hours, and 24 hours of hypoxia. Blots are representative of three independent experiments in each condition; α-tubulin was used as the loading control. ( B ) Representative immunoblots of HIF1α protein from retinas of C57BL/6J mice at 1 dprd and 3 dprd. Blots are representative of three independent animals in each condition; α-tubulin was used as the loading control. ( C ) Relative mRNA expression of Hspa5 obtained from retinas of Rho/Cre + and HIF1αΔrod mice at 3 dprd compared to the contralateral eye (fellow). Pum-1 was used as a housekeeping control gene for normalization. ( D ) Relative mRNA expression of Ddit3 obtained from retinas of Rho/Cre + and HIF1αΔrod mice at 3 dprd compared to the contralateral eye (fellow). Pum-1 was used as a housekeeping control gene for normalization. ( E ) Relative mRNA expression of Pgc1a and Tfam obtained from retinas of Rho/Cre + and HIF1αΔrod mice at 3 dprd compared to the contralateral eye (fellow). Pum-1 was used as a housekeeping control gene for normalization. ( F ) Representative immunoblots of Parkin and FUNDC1 proteins from retinas of Rho/Cre + and HIF1αΔrod mice at 3 dprd compared to the contralateral eye (fellow). GAPDH and α-tubulin were used as the loading control. ( G ) Relative mRNA expression of Prkn obtained from retinas of Rho/Cre+ and HIF1αΔrod mice at 3 dprd compared to the contralateral eye (fellow). Pum-1 was used as a housekeeping control gene for normalization. ( H ) Relative mRNA expression of Pink obtained from retinas of Rho/Cre + and HIF1αΔrod mice at 3 dprd compared to the contralateral eye (fellow). Pum-1 was used as a housekeeping control gene for normalization. Bar graphs represent mean ± SEM. Statistical analysis was performed using two-way ANOVA with repeated measures followed by Tukey's test. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: After fixation, the cells were permeabilized with 0.1% Triton X-100, blocked using 2% BSA, and subsequently subjected to an overnight incubation with the antibodies against FUNDC1 (1:250; Novus Biologicals) or anti-mouse LC3B (#83506, 1:250; Cell Signaling Technology).

Techniques: Western Blot, Control, Expressing

Journal: The American journal of pathology

Article Title: Molecular and functional alterations in a mouse cardiac model of Friedreich ataxia: activation of the integrated stress response, eIF2α phosphorylation, and the induction of downstream targets.

doi: 10.1016/j.ajpath.2013.05.032

Figure Lengend Snippet: Figure 6 Proposed pathogenesis of heart failure in FA. Frataxin defi- ciency results in the early activation of the ISR via phosphorylation (P) of eIF2a and induction of its downstream stress-inducible molecules, including Atf4 and Chop. The precise stress stimulus for the ISR activation is unclear, but may involve early and pronounced perturbation in heme synthesis (markedly reduced Fech expression) acting through Hri in KO mice, compared to WT mice. In addition, the later involvement of ER stress (up-regulation of Bip and cleaved caspase-12) could potentiate the prolonged activation of the ISR pathway. ISR activation subsequently enhances autophagy (up- regulation of Atg3, LC3-II, p62, and Fundc1) and apoptosis (increased expression of Chop, Bax and Bcl-2, and cleaved caspase-12), leading to cardiomyocyte death. Histological alterations are first apparent from 5 weeks of age with iron accumulation, followed by fibrosis and cardiomyocyte degeneration from 6 weeks of age. Severe progressive reduction in cardiac function ensues, resulting in heart failure. Casp12, caspase-12.

Article Snippet: Protein isolation and Western blot analysis were performed using established techniques.3 The primary antibodies used were against TfR1 (13-6890; Life Technologies), Atf4 (sc-200; SantaCruzBiotechnology,SantaCruz,CA),Mthfd2 (ab37840; Abcam, Cambridge, UK), Asns (1732-1; Epitomics, Burlingame,CA), Chop (SC-575; Santa Cruz), p-eIF2a (Ser51; 3398; Cell Signaling Technology, Danvers, MA), eIF2a (2103; Cell Signaling Technology), Atg3 (3415; Cell Signaling Technology), LC3 (PD014; MBL International, Woburn, MA), p62 (GP62-C; Progen Biotechnik, Heidelberg, Germany), caspase12 (2202; Cell Signaling Technology), Bax (2772; Cell Signaling Technology), Bcl-2 (2870; Cell Signaling Technology), Itgb1bp3 (K0099-3; MBL International), p-Perk (Thr980; 3179; Cell Signaling Technology), Perk (3192; Cell Signaling Technology), Bip (3177; Cell Signaling Technology), Hri (07-728; Millipore, Sydney, Australia), Fech (from Harry Dailey, University of Georgia30), Fundc1 (ARP53280_P050; Aviva Systems Biology, San Diego, CA), and Gapdh (SC-25778; Santa Cruz Biotechnology).

Techniques: Activation Assay, Phospho-proteomics, Expressing

Journal: Nature Communications

Article Title: Endothelial FUNDC1 regulates metabolic reprogramming and the obesity-diabetes transition through the SIRT3/GATA2/endothelin-1 axis

doi: 10.1038/s41467-026-68548-4

Figure Lengend Snippet: a – c CD31 positive ECs (CD31 + ECs) were isolated from white adipose tissue (WAT) of mice fed with indicated durations (2, 4, 6 months) of HFD and subjected to western blot analysis ( n = 4 mice/group) or Q-PCR analysis ( n = 6 mice/group). d – f HUVECs were incubated with 200 μM PA conjugated to fatty acid-free BSA, and culture medium containing PA or BSA was refreshed daily throughout the 7-day stimulation period to maintain a consistent lipid challenge. Cells were then subjected to western blot analysis or Q-PCR analysis. n = 3 independent experiments. g CD31 positive cells were removed from the SVF of Fundc1 f/Y Cdh5 - or Fundc1 f/Y Cdh5 + mice (CD31 - SVF). Normal SVF containing CD31 positive cells was used as control (CD31 + SVF). Cells were then differentiated into adipocytes. Scale bar: 50 µm. h Intracellular TG content in ( g ). n = 6 independent experiments. i Insulin-stimulated 2-NBDG uptake in primary adipocytes isolated from Fundc1 f/Y Cdh5 - or Fundc1 f/Y Cdh5 + mice in the present of bovine serum albumin (BSA) or palmitic acid (PA). n = 8 independent experiments. Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, n.s. p > 0.05.

Article Snippet: The aortic sections were incubated with FUNDC1 (Biorbyt, orb156964), 8-OHdG (abcam, ab62623), or ET-1 (Novus, NB300-526) antibody at 4 °C overnight and followed by incubation with EnVision® + Dual Link System-HRP (Dako Cytomation, 3468) and the signal was visualized using the Liquid DAB+ Substrate Chromogen System and imaged using an inverted phase contrast microscope (Nikon, Tokyo, Japan).

Techniques: Isolation, Western Blot, Incubation, Control

Journal: Nature Communications

Article Title: Endothelial FUNDC1 regulates metabolic reprogramming and the obesity-diabetes transition through the SIRT3/GATA2/endothelin-1 axis

doi: 10.1038/s41467-026-68548-4

Figure Lengend Snippet: a – l Fundc1 f/Y Cdh5 + mice and littermates ( Fundc1 f/Y Cdh5 - ) were kept on HFD for 6 months. a Changes in body weight during HFD feeding. * p < 0.05, ** p < 0.01 (HFD- Fundc1 f/Y Cdh5 - vs . HFD- Fundc1 f/Y Cdh5 +) , n = 15–20. b Fat percentage of mice. n = 15–20 mice/group. c Adipose tissue weight/body weight. n = 15–20 mice/group. d Representative images of H&E staining of SubWAT. e Quantification of siz e s and numbers of adipocytes under HFD feeding. n = 15–20 mice/group. f Quantification of adipocyte diameter. n = 15–20 mice/group. g Glucose tolerance test (GTT) and quantification of the area under curve, n = 7-8 mice/group. h Insulin tolerance test (ITT) and quantification of the area under curve, n = 7–8 mice/group. i , j Whole body respiratory exchange ratio (RER; VCO 2 /VO 2 , n = 8 mice/group). k , l Energy expenditure normalized to body weight, n = 8 mice/group. Data are presented as mea n ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, n.s. p > 0.05.

Article Snippet: The aortic sections were incubated with FUNDC1 (Biorbyt, orb156964), 8-OHdG (abcam, ab62623), or ET-1 (Novus, NB300-526) antibody at 4 °C overnight and followed by incubation with EnVision® + Dual Link System-HRP (Dako Cytomation, 3468) and the signal was visualized using the Liquid DAB+ Substrate Chromogen System and imaged using an inverted phase contrast microscope (Nikon, Tokyo, Japan).

Techniques: Staining

Journal: Nature Communications

Article Title: Endothelial FUNDC1 regulates metabolic reprogramming and the obesity-diabetes transition through the SIRT3/GATA2/endothelin-1 axis

doi: 10.1038/s41467-026-68548-4

Figure Lengend Snippet: Mice were placed on a high-fat diet (HFD) and infused with ET-1 (10 µg/kg/day) or saline concurrently, starting at the onset of HFD feeding, using subcutaneously implanted osmotic pumps for 4 weeks. a Representative pictures of whole body, epididymal white adipose tissue (EpiWAT), subcutaneous white adipose tissue (SubWAT), and brown adipose tissue (BAT). b Changes in body weight during HFD feeding periods (left panel), * p < 0.05, ** p < 0.01 (Saline- Fundc1 f/Y Cdh5 - vs . Saline- Fundc1 f/Y Cdh5 + ), # p < 0.05, ## p < 0.01 (ET-1- Fundc1 f/Y Cdh5 - vs . ET-1- Fundc1 f/Y Cdh5 + ), and relative weights of adipose tissues ( right panel). n = 7–8 mice/group. c Glucose tolerance test (IPGTT) and insulin tolerance test (ITT). n = 7–8 mice/group. d Fasting blood glucose (GLU) level. n = 7–8 mice/group. e Serum insulin level. n = 7–8 mice/group. f Serum free fatty acid (FFA) level. n = 7–8 mice/group. g – i Aortic ring vasoreactivity assay in ET-1-infused mice. n = 6 mice/group. j , k Circumferential cyclic strain ( j ) and (PWV) pulse wave velocity ( k ) of carotid artery. n = 6 mice/group. Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, n.s. p > 0.05.

Article Snippet: The aortic sections were incubated with FUNDC1 (Biorbyt, orb156964), 8-OHdG (abcam, ab62623), or ET-1 (Novus, NB300-526) antibody at 4 °C overnight and followed by incubation with EnVision® + Dual Link System-HRP (Dako Cytomation, 3468) and the signal was visualized using the Liquid DAB+ Substrate Chromogen System and imaged using an inverted phase contrast microscope (Nikon, Tokyo, Japan).

Techniques: Saline

Journal: Nature Communications

Article Title: Endothelial FUNDC1 regulates metabolic reprogramming and the obesity-diabetes transition through the SIRT3/GATA2/endothelin-1 axis

doi: 10.1038/s41467-026-68548-4

Figure Lengend Snippet: a HUVECs were transfected with control siRNA (siCtrl) or FUNDC1 siRNA ( siFUNDC1 ) and subjected to immunoblotting analysis. b Quantification of protein levels of ECE1, GATA2, and AP1. n = 6 independent experiments. c HUVECs were transfected with control siRNA (siCtrl) or FUNDC1 siRNA ( siFUNDC1 ) and subjected to Q-PCR analysis to detect ECE1, AP1, and JUN mRNA levels. n = 8 independent experiments. d HUVECs were transfected with control Myc-Flag-tagged plasmid ( Myc-Flag-Ctrl ) or Myc-Flag-FUNDC1 plasmid ( Myc-Flag-FUNDC1 ) and subjected to immunoblotting analysis. e Quantification of protein levels of FUNDC1, ECE1, GATA2 and AP1 in ( d ). n = 6 independent experiments. f HUVECs were transfected with control siRNA (siCtrl) or GATA2 siRNA ( siGATA2 ) and subjected to Q-PCR analysis to detect EDN1 mRNA levels. n = 8 independent experiments. g HUVECs were transfected with control siRNA (siCtrl) or FUNDC1 siRNA ( siFUNDC1 ) and were then treat with 200 µM PA for indicated time periods. h - j Quantification of protein levels of FUNDC1, GATA2, and ET-1 in ( g ). n = 4 independent experiments. Data are presented as mean ± SD, * p < 0.05, ** p < 0.01, *** p < 0.001, n.s. p > 0.05.

Article Snippet: The aortic sections were incubated with FUNDC1 (Biorbyt, orb156964), 8-OHdG (abcam, ab62623), or ET-1 (Novus, NB300-526) antibody at 4 °C overnight and followed by incubation with EnVision® + Dual Link System-HRP (Dako Cytomation, 3468) and the signal was visualized using the Liquid DAB+ Substrate Chromogen System and imaged using an inverted phase contrast microscope (Nikon, Tokyo, Japan).

Techniques: Transfection, Control, Western Blot, Plasmid Preparation

Journal: Nature Communications

Article Title: Endothelial FUNDC1 regulates metabolic reprogramming and the obesity-diabetes transition through the SIRT3/GATA2/endothelin-1 axis

doi: 10.1038/s41467-026-68548-4

Figure Lengend Snippet: a HUVECs were transfected with control siRNA (siCtrl) or SIRT3 siRNA ( siSIRT3 ) and subjected to immunoblotting analysis. b Quantification of protein levels in ( a ). n = 8–10 independent experiments. c HUVECs were transfected with control siRNA (siCtrl) or SIRT3 siRNA ( siSIRT3 ) and subjected to Q-PCR analysis to detect mRNA levels of GATA2 and EDN1 . n = 8 independent experiments. d The interactions between SIRT3 and FUNDC1 or GATA2 in HUVECs were determined by immunoprecipitation (IP) and immunoblotting (IB) analysis. e The co-localization of SIRT3 and FUNDC1 or GATA2 were determined by immunofluorescence staining. Nuclei were stained by DAPI. MERGE1 indicates merge of SIRT3 and DAPI; MERGE2 indicates merge of FUNDC1, SIRT3, and DAPI; MERGE3 indicates merge of GATA2 and DAPI; MERGE4 indicates merge of GATA2, SIRT3, and DAPI. Scale bar: 10 µm. f HUVECs were transfected with control siRNA (siCtrl) or FUNDC1 siRNA ( siFUNDC1 ) in the presence of BSA or 200 µM PA and subjected to immunofluorescence staining analysis. MERGE1 indicates merge of SIRT3 and DAPI; MERGE2 indicates merge of FUNDC1, SIRT3, and DAPI. Scale bar: 10 µm. g Pearson’s coefficient calculated by ZEN software, indicates the co-localization of FUNDC1 and SIRT3. n = 9 cells from three independent experiments. h Quantification of nuclear SIRT3 level. i HUVECs were transfected with control siRNA (siCtrl) or FUNDC1 siRNA ( siFUN ) in the presence of BSA or 200 µM PA. Non-nuclear fraction (Non-Nuc), nuclear fraction (Nuc), and mitochondria fraction (Mito) were prepared and subjected to immunoblotting analysis. n = 9 cells from three independent experiments. j Quantification of protein level of SIRT3 long isoform (SIRT3-L) in the nucleus (Nuclear SIRT3-L) and in mitochondria (Mitochondrial SIRT3-L). n = 4 independent experiments. k HUVECs were double transfected with control siRNA (siCtrl) or FUNDC1 ( siFUN ) and SIRT3 siRNA and subjected to immunoblotting analysis. l Quantification of GATA2 protein level in ( k ). n = 5 independent experiments. m HUVECs were double transfected with either control siRNA (siCtrl) or FUNDC1 and SIRT3 siRNA and subjected to Q-PCR analysis to detect END1 mRNA level. n = 8 independent experiments. Data are presented as mean ± SD, * p < 0.05, ** p < 0.01, *** p < 0.001, n.s. p > 0.05.

Article Snippet: The aortic sections were incubated with FUNDC1 (Biorbyt, orb156964), 8-OHdG (abcam, ab62623), or ET-1 (Novus, NB300-526) antibody at 4 °C overnight and followed by incubation with EnVision® + Dual Link System-HRP (Dako Cytomation, 3468) and the signal was visualized using the Liquid DAB+ Substrate Chromogen System and imaged using an inverted phase contrast microscope (Nikon, Tokyo, Japan).

Techniques: Transfection, Control, Western Blot, Immunoprecipitation, Immunofluorescence, Staining, Software

Journal: Nature Communications

Article Title: Endothelial FUNDC1 regulates metabolic reprogramming and the obesity-diabetes transition through the SIRT3/GATA2/endothelin-1 axis

doi: 10.1038/s41467-026-68548-4

Figure Lengend Snippet: a HUVECs were transfected with control siRNA (siCtrl) or SIRT3 siRNA and then treated with 50 µg/ml cycloheximide (CHX) for 240 min. Cell lysates were subjected to immunoblotting analysis. b Quantification of GATA2 protein level in ( a ). n = 4 independent experiments. c Acetylated GATA2 level was determined by IP of acetylate lysine (Ac-lysine) in cells transfected with SIRT3 siRNA and followed by immunoblotting (IB) of GATA2. d Quantification of the enrichment of acetylated GATA2 in ( c ). n = 4 independent experiments. e Acetylated GATA2 level was determined by IP of Ac-lysine in cells treated with PA (200 µM) and followed by immunoblotting (IB) GATA2. f Quantification of the enrichment of acetylated GATA2 in ( e ). n = 4 independent experiments. g HUVECs were transfected with control siRNA (siCtrl) or SIRT3 siRNA ( siSIRT3 ) and subjected to immunoblotting analysis. h HUVECs were transfected with control Myc-Flag-tagged plasmid ( Myc-Flag-Ctrl ) or Myc-Flag-SIRT3 plasmid ( Myc-Flag-SIRT3 ) and subjected to immunoblotting analysis. i , j Quantification of protein levels of FUNDC1. n = 6–9 independent experiments. k HUVECs were transfected with control siRNA (siCtrl) or SIRT3 siRNA ( siSIRT3 and subjected to Q-PCR analysis to detect FUNDC1 mRNA level. n = 8. l HUVECs were transfected with control siRNA (siCtrl) or SIRT3 siRNA and then treated with 50 µg/ml cycloheximide (CHX) for 240 min. Cell lysates were subjected to immunoblotting analysis. m Quantification of GATA2 protein level in ( l ). n = 3 independent experiments. Data are presented as mean ± SD, in ( b and m ), star represents the comparison between groups siCtrl and siSIRT3 at the same treatment intervals, pound represents the comparison with control group (time = 0 min). * p < 0.05, ** p < 0.01, *** p < 0.001, n.s. p > 0.05. # p < 0.05, ## p < 0.01, ### p < 0.001, n.s. p > 0.05.

Article Snippet: The aortic sections were incubated with FUNDC1 (Biorbyt, orb156964), 8-OHdG (abcam, ab62623), or ET-1 (Novus, NB300-526) antibody at 4 °C overnight and followed by incubation with EnVision® + Dual Link System-HRP (Dako Cytomation, 3468) and the signal was visualized using the Liquid DAB+ Substrate Chromogen System and imaged using an inverted phase contrast microscope (Nikon, Tokyo, Japan).

Techniques: Transfection, Control, Western Blot, Plasmid Preparation, Comparison

Journal: Nature Communications

Article Title: Endothelial FUNDC1 regulates metabolic reprogramming and the obesity-diabetes transition through the SIRT3/GATA2/endothelin-1 axis

doi: 10.1038/s41467-026-68548-4

Figure Lengend Snippet: a Potential KFERQ-like motifs of human FUNDC1 and SIRT3. b Interactions between HSC70 with FUNDC1 and SIRT3-L in HUVECs were detected by IP and IB. c Co-staining of FUNDC1 with HSC70 and SIRT3-L by IF. d Interaction of SIRT3-L and FUNDC1 was analyzed in HUVECs transfected with HSC70 siRNA. e Quantification of protein levels in HUVECS transfected with HSC70 siRNA. f Quantification of the binding activity of FUNDC1 with SIRT3-L in HSC70 knockdown cells, indicated by the enrichment of FUNDC1. n = 4 independent experiments. g HUVECs were transfected with control siRNA (siCtrl) or HSC70 siRNA ( siHSC70 ) in the presence of BSA or 200 µM PA. Non-nuclear fraction (Non-Nuc), nuclear fraction (Nuc), and mitochondria fraction (Mito) were prepared and subjected to immunoblotting analysis. h Quantification of protein level of SIRT3 long isoform (SIRT3-L) in the nucleus (Nuclear SIRT3-L) and in mitochondria (Mitochondrial SIRT3-L). n = 4 independent experiments. Data are presented as mean ± SD, * p < 0.05, ** p < 0.01 derived from Student’s t tests.

Article Snippet: The aortic sections were incubated with FUNDC1 (Biorbyt, orb156964), 8-OHdG (abcam, ab62623), or ET-1 (Novus, NB300-526) antibody at 4 °C overnight and followed by incubation with EnVision® + Dual Link System-HRP (Dako Cytomation, 3468) and the signal was visualized using the Liquid DAB+ Substrate Chromogen System and imaged using an inverted phase contrast microscope (Nikon, Tokyo, Japan).

Techniques: Staining, Transfection, Binding Assay, Activity Assay, Knockdown, Control, Western Blot, Derivative Assay

Journal: Nature Communications

Article Title: Endothelial FUNDC1 regulates metabolic reprogramming and the obesity-diabetes transition through the SIRT3/GATA2/endothelin-1 axis

doi: 10.1038/s41467-026-68548-4

Figure Lengend Snippet: Plasma and endothelium were collected from age-matched healthy donors ( n = 17) or donors with both obesity and T2DM (Obesity&T2DM, n = 19). a Characteristics of human donors. Age, body mass index (BMI), and glycohemoglobin A1c (HbA1c) levels are represented as mean ± SD. b Plasma ET-1 level in human donors. n = 17–19 donors/group. c Spearman’s correlation of plasma ET-1 level and BMI. d Spearman’s correlation of plasma ET-1 level and HbA1c. e qPCR analysis of FUNDC1 , END1 and GATA2 mRNA levels in the intima isolated from human small arterioles. n = 17–19 donors/group. f Spearman’s correlation of mRNA levels of FUNDC1 and END1 . g Spearman’s correlation of mRNA levels of GATA2 and END1 . h Spearman’s correlation of intimal FUNDC1 mRNA and plasma ET-1. i , j Human small arterioles were collected from age-matched healthy donors or donors with both obesity and T2DM (Obesity&T2DM) and subjected to western blot analysis ( i and j ). n = 6 donors/group. k , l Human femoral arteries were collected from age-matched healthy donors (n = 7) or donors with both obesity and T2DM (Obesity&T2DM; n = 5) and subjected to IHC staining. Scale bar: 50 µm. Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, n.s. p > 0.05.

Article Snippet: The aortic sections were incubated with FUNDC1 (Biorbyt, orb156964), 8-OHdG (abcam, ab62623), or ET-1 (Novus, NB300-526) antibody at 4 °C overnight and followed by incubation with EnVision® + Dual Link System-HRP (Dako Cytomation, 3468) and the signal was visualized using the Liquid DAB+ Substrate Chromogen System and imaged using an inverted phase contrast microscope (Nikon, Tokyo, Japan).

Techniques: Clinical Proteomics, Isolation, Western Blot, Immunohistochemistry

Journal: Nature Communications

Article Title: Endothelial FUNDC1 regulates metabolic reprogramming and the obesity-diabetes transition through the SIRT3/GATA2/endothelin-1 axis

doi: 10.1038/s41467-026-68548-4

Figure Lengend Snippet: In the healthy state, endothelial FUNDC1 anchors SIRT3-L in mitochondria, limiting its nuclear accumulation and regulating its interaction with GATA2, thereby maintaining normal ET-1 production and preserving vascular and metabolic homeostasis. Under overnutrition-induced metabolic stress, SIRT3-L is released from GATA2, contributing to enhanced GATA2-mediated ET-1 transcription. Elevated ET-1 promote angiogenesis to provide the space for adipocyte hyperplasia at the early stage and later exacerbate insulin resistance and thus driving the progression from obesity to T2DM. Importantly, SIRT3-L translocation to mitochondria also facilitates FUNDC1 degradation at the early obese stage, which triggers a compensatory increase in FUNDC1 transcription and ultimately results in FUNDC1 overexpression in a late-stage. The elevation of FUNDC1 further enhances mitochondrial recruitment of SIRT3-L, amplifying ET-1 production. In contrast, in Fundc1 EC-specific knockout (ECKO) mice, the absence of FUNDC1 prevents mitochondrial recruitment of SIRT3-L, leading to its nuclear retention, where SIRT3-L promotes GATA2 degradation and suppresses ET-1 transcription. The loss of Fundc1 in ECs not only suppresses angiogenesis at early overnutrition stage but also prevents EC senescence at late stage to attenuate the potential diabetic vascular complications.

Article Snippet: The aortic sections were incubated with FUNDC1 (Biorbyt, orb156964), 8-OHdG (abcam, ab62623), or ET-1 (Novus, NB300-526) antibody at 4 °C overnight and followed by incubation with EnVision® + Dual Link System-HRP (Dako Cytomation, 3468) and the signal was visualized using the Liquid DAB+ Substrate Chromogen System and imaged using an inverted phase contrast microscope (Nikon, Tokyo, Japan).

Techniques: Preserving, Translocation Assay, Over Expression, Knock-Out

Journal: Redox Biology

Article Title: A novel role of KEAP1/PGAM5 complex: ROS sensor for inducing mitophagy

doi: 10.1016/j.redox.2021.102186

Figure Lengend Snippet:

Article Snippet: FUNDC1 shRNA , Origene , Cat# TR702819.

Techniques: Recombinant, Lysis, Extraction, Blocking Assay, Isolation, DC Protein Assay, Caspase-Glo Assay, shRNA, Generated, Plasmid Preparation, Synthesized, Variant Assay

Journal: Open Medicine

Article Title: Methylation regulation for FUNDC1 stability in childhood leukemia was up-regulated and facilitates metastasis and reduces ferroptosis of leukemia through mitochondrial damage by FBXL2

doi: 10.1515/med-2023-0810

Figure Lengend Snippet: Expression levels of FUNDC1 in CL. FUNDC1 mRNA expression (a) and (b), ROC (c) in patients with CL; FUNDC1 mRNA and protein expression (d) and (e) in leukemia cells. Normal, normal serum sample; Cancer, patients with CL; ** p < 0.01 compared with normal serum sample or HS-5 cells.

Article Snippet: FUNDC1 plasmids (sc-428195, Santa Cruz Biotechnology, Inc.) or si-FUNDC1 plasmids (sc-145273, Santa Cruz Biotechnology, Inc.) were transfected into GC cell lines using Lipofectamine 2000.

Techniques: Expressing

Journal: Open Medicine

Article Title: Methylation regulation for FUNDC1 stability in childhood leukemia was up-regulated and facilitates metastasis and reduces ferroptosis of leukemia through mitochondrial damage by FBXL2

doi: 10.1515/med-2023-0810

Figure Lengend Snippet: FUNDC1 up-regulation promotes leukemia metastasis. FUNDC1 mRNA expression (a), cell growth (b), metastasis (c), and EDU assay (d) in vitro model of leukemia by FUNDC1 up-regulation; FUNDC1 mRNA expression (e), cell growth (f), metastasis (g), and EDU assay (h) in vitro model of leukemia by FUNDC1 down-regulation. (i). Fluorescence images showing reduced proliferation in leukemia cells with FUNDC1 knockdown, as evidenced by fewer EDU-positive cells. (j). Micrographs illustrating decreased migration of leukemia cells after FUNDC1 knockdown. Vector, negative control group; FUNDC1, over-expression of FUNDC1 group; Si-nc, si-negative control group; Si-FUNDC1, down-regulation of FUNDC1 group; ** p < 0.01 compared with negative control group or si-negative control group.

Article Snippet: FUNDC1 plasmids (sc-428195, Santa Cruz Biotechnology, Inc.) or si-FUNDC1 plasmids (sc-145273, Santa Cruz Biotechnology, Inc.) were transfected into GC cell lines using Lipofectamine 2000.

Techniques: Expressing, EdU Assay, In Vitro, Fluorescence, Knockdown, Migration, Plasmid Preparation, Negative Control, Over Expression

Journal: Open Medicine

Article Title: Methylation regulation for FUNDC1 stability in childhood leukemia was up-regulated and facilitates metastasis and reduces ferroptosis of leukemia through mitochondrial damage by FBXL2

doi: 10.1515/med-2023-0810

Figure Lengend Snippet: FUNDC1 up-regulation reduces ferroptosis by inducing mitochondrial damage. JC-1 levels (a), MPT (b), mitochondrial damage (c), PI-positive cells (d), iron content (e), caspase-3/9 activity (f) and (g), LDH activity levels (h), GSH levels (i), and GPX4 protein expression (j) in vitro model of leukemia by FUNDC1 up-regulation; JC-1 levels (k), MPT (i), mitochondrial damage (l), mitochondrial damage (m), PI-positive cells (n), iron content (o), caspase-3/9 activity (p) and (q), LDH activity levels (r), GSH levels (s), and GPX4 protein expression (t) in vitro model of leukemia by FUNDC1 down-regulation. Vector, negative control group; FUNDC1, over-expression of FUNDC1 group; Si-nc, si-negative control group; Si-FUNDC1, down-regulation of FUNDC1 group; ** p < 0.01 compared with negative control group or si-negative control group.

Article Snippet: FUNDC1 plasmids (sc-428195, Santa Cruz Biotechnology, Inc.) or si-FUNDC1 plasmids (sc-145273, Santa Cruz Biotechnology, Inc.) were transfected into GC cell lines using Lipofectamine 2000.

Techniques: Activity Assay, Expressing, In Vitro, Plasmid Preparation, Negative Control, Over Expression

Journal: Open Medicine

Article Title: Methylation regulation for FUNDC1 stability in childhood leukemia was up-regulated and facilitates metastasis and reduces ferroptosis of leukemia through mitochondrial damage by FBXL2

doi: 10.1515/med-2023-0810

Figure Lengend Snippet: FUNDC1 up-regulation induces FBXL2 expression in a leukemia model. FBXL2 mRNA expression (a), FBXL2/FUNDC1 protein expression (b) and (c), FBXL2/FUNDC1 expression confocal microscope (d), FBXL2 protein interlinked with FUNDC1 protein (e), FBXL2 ubiquitination (f). Vector, negative control group; FUNDC1, over-expression of FUNDC1 group; Si-nc, si-negative control group; Si-FUNDC1, down-regulation of FUNDC1 group; ** p < 0.01 compared with negative control group or si-negative control group.

Article Snippet: FUNDC1 plasmids (sc-428195, Santa Cruz Biotechnology, Inc.) or si-FUNDC1 plasmids (sc-145273, Santa Cruz Biotechnology, Inc.) were transfected into GC cell lines using Lipofectamine 2000.

Techniques: Expressing, Microscopy, Ubiquitin Proteomics, Plasmid Preparation, Negative Control, Over Expression

Journal: Open Medicine

Article Title: Methylation regulation for FUNDC1 stability in childhood leukemia was up-regulated and facilitates metastasis and reduces ferroptosis of leukemia through mitochondrial damage by FBXL2

doi: 10.1515/med-2023-0810

Figure Lengend Snippet: Mitochondrial damage regulates the effects of FUNDC1 up-regulation on ferroptosis in leukemia. JC-1 levels (a), MPT (b), mitochondrial damage (c), PI-positive cells (d), LDH activity levels (e), iron content (f), GSH levels (g), and GPX4 protein expression (h) in vitro model of leukemia by FUNDC1 up-regulation and activator; JC-1 levels (i), MPT (j), mitochondrial damage (k), PI-positive cells (l), LDH activity levels (m), iron content (n), GSH levels (o), and GPX4 protein expression (p) in vitro model of leukemia by FUNDC1 down-regulation and inhibitor. Vector, negative control group; FUNDC1, over-expression of FUNDC1 group; Si-nc, si-negative control group; si-FUNDC1, down-regulation of FUNDC1 group; ** p < 0.01 compared with negative control group or si-negative control group; ## p < 0.01 compared with FUNDC1 or si-FUNDC1 group.

Article Snippet: FUNDC1 plasmids (sc-428195, Santa Cruz Biotechnology, Inc.) or si-FUNDC1 plasmids (sc-145273, Santa Cruz Biotechnology, Inc.) were transfected into GC cell lines using Lipofectamine 2000.

Techniques: Activity Assay, Expressing, In Vitro, Plasmid Preparation, Negative Control, Over Expression

Journal: Open Medicine

Article Title: Methylation regulation for FUNDC1 stability in childhood leukemia was up-regulated and facilitates metastasis and reduces ferroptosis of leukemia through mitochondrial damage by FBXL2

doi: 10.1515/med-2023-0810

Figure Lengend Snippet: Mitochondrial damage regulates the effects of FUNDC1 up-regulation on metastasis in leukemia. Cell growth (a), metastasis (b), and EDU assay (c) in vitro model of leukemia by FUNDC1 up-regulation; cell growth (d), metastasis (e), and EDU assay (f) in vitro model of leukemia by FUNDC1 down-regulation. Vector, negative control group; FUNDC1, over-expression of FUNDC1 group; Si-nc, si-negative control group; Si-FUNDC1, down-regulation of FUNDC1 group; ** p < 0.01 compared with negative control group or si-negative control group.

Article Snippet: FUNDC1 plasmids (sc-428195, Santa Cruz Biotechnology, Inc.) or si-FUNDC1 plasmids (sc-145273, Santa Cruz Biotechnology, Inc.) were transfected into GC cell lines using Lipofectamine 2000.

Techniques: EdU Assay, In Vitro, Plasmid Preparation, Negative Control, Over Expression

Journal: Open Medicine

Article Title: Methylation regulation for FUNDC1 stability in childhood leukemia was up-regulated and facilitates metastasis and reduces ferroptosis of leukemia through mitochondrial damage by FBXL2

doi: 10.1515/med-2023-0810

Figure Lengend Snippet: Methylation controls FUNDC1 stability. m6A modification site of FUNDC1 (a), METTL3-mediated FUNDC1 m6A modifications (b) and (c), the position of m6A motifs within FUNDC1 transcript sequence (d), luciferase reporter activity level (e), m6A levels of FUNDC1 (f) and (g), ** p < 0.01 compared with vector or negative or IgG group.

Article Snippet: FUNDC1 plasmids (sc-428195, Santa Cruz Biotechnology, Inc.) or si-FUNDC1 plasmids (sc-145273, Santa Cruz Biotechnology, Inc.) were transfected into GC cell lines using Lipofectamine 2000.

Techniques: Methylation, Modification, Sequencing, Luciferase, Activity Assay, Plasmid Preparation