Journal: Frontiers in Cardiovascular Medicine

Article Title: Herb pair of Astragali Radix-Descurainiae Semen attenuate heart failure through the myosin VI-Tom1 complex mediated autophagy

doi: 10.3389/fcvm.2025.1599746

Figure Lengend Snippet: Relative protein expression and statistics of MYO6 and Tom1 in rat myocardium in each group (A,B) . Relative gene expression and statistics of MYO6 and Tom1 in rat myocardium in each group (C) Multiple immunofluorescence staining images of rat myocardium in the Control, ISO, and AR-DS-H groups. MYO6 and Tom1 co-expressing cells were labeled using white arrows. (60×, scale bar = 20 μm) (D) Analysis of the average fluorescence intensity of MYO6 and Tom1 in the myocardium of rats from the Control, ISO, and AR-DS-H groups (E) Compared to the Control group: ** P < 0.01, * P < 0.05; compared to the ISO group: ## P < 0.01, # P < 0.05, n = 3. Protein docking image of MYO6 and Tom1, where green represents the MYO6 protein and sky blue represents the Tom1 protein (F)

Article Snippet: Primary antibodies including GAPDH (1:10,000, AC001, ABclonal Technology, Hubei, China)), LC3 (1:1,000, ab192890, Abcam, UK), p62 (1:1,000, ab109012, Abcam, UK), Beclin1 (1:1,000, 11306-1-AP, Proteintech, Wuhan, China), MYO6 (1:500, 26778-1-AP, Proteintech, Wuhan, China), and Tom1 (1:500, sc-514430, Santa Cruz Biotechnology, USA) were added and incubated overnight at 4°C.

Techniques: Expressing, Gene Expression, Immunofluorescence, Staining, Control, Labeling, Fluorescence

Journal: Frontiers in Cardiovascular Medicine

Article Title: Herb pair of Astragali Radix-Descurainiae Semen attenuate heart failure through the myosin VI-Tom1 complex mediated autophagy

doi: 10.3389/fcvm.2025.1599746

Figure Lengend Snippet: Mode of action, 3D, active pocket, and 2D maps of 10 compounds in MYO6-Tom1 complex and AR-DS decoction. The compounds were Ononin (A) , Astragaloside-IV (B) , Rutin (C) , Folic-acid (D) , Daidzein (E) , Isorhamnetin (F) , Quercetin (G) , Beta-Sitosterol (H) , Kaempferol (I) , Formononetin (J) .

Article Snippet: Primary antibodies including GAPDH (1:10,000, AC001, ABclonal Technology, Hubei, China)), LC3 (1:1,000, ab192890, Abcam, UK), p62 (1:1,000, ab109012, Abcam, UK), Beclin1 (1:1,000, 11306-1-AP, Proteintech, Wuhan, China), MYO6 (1:500, 26778-1-AP, Proteintech, Wuhan, China), and Tom1 (1:500, sc-514430, Santa Cruz Biotechnology, USA) were added and incubated overnight at 4°C.

Techniques:

Journal: Frontiers in Cardiovascular Medicine

Article Title: Herb pair of Astragali Radix-Descurainiae Semen attenuate heart failure through the myosin VI-Tom1 complex mediated autophagy

doi: 10.3389/fcvm.2025.1599746

Figure Lengend Snippet: AR-DS may regulate myocardial autophagy in HF rats through the MYO6-Tom1 complex. (Created using Figdraw ).

Article Snippet: Primary antibodies including GAPDH (1:10,000, AC001, ABclonal Technology, Hubei, China)), LC3 (1:1,000, ab192890, Abcam, UK), p62 (1:1,000, ab109012, Abcam, UK), Beclin1 (1:1,000, 11306-1-AP, Proteintech, Wuhan, China), MYO6 (1:500, 26778-1-AP, Proteintech, Wuhan, China), and Tom1 (1:500, sc-514430, Santa Cruz Biotechnology, USA) were added and incubated overnight at 4°C.

Techniques:

Journal: Frontiers in Cardiovascular Medicine

Article Title: Herb pair of Astragali Radix-Descurainiae Semen attenuate heart failure through the myosin VI-Tom1 complex mediated autophagy

doi: 10.3389/fcvm.2025.1599746

Figure Lengend Snippet: Relative protein expression and statistics of MYO6 and Tom1 in rat myocardium in each group (A,B) . Relative gene expression and statistics of MYO6 and Tom1 in rat myocardium in each group (C) Multiple immunofluorescence staining images of rat myocardium in the Control, ISO, and AR-DS-H groups. MYO6 and Tom1 co-expressing cells were labeled using white arrows. (60×, scale bar = 20 μm) (D) Analysis of the average fluorescence intensity of MYO6 and Tom1 in the myocardium of rats from the Control, ISO, and AR-DS-H groups (E) Compared to the Control group: ** P < 0.01, * P < 0.05; compared to the ISO group: ## P < 0.01, # P < 0.05, n = 3. Protein docking image of MYO6 and Tom1, where green represents the MYO6 protein and sky blue represents the Tom1 protein (F)

Article Snippet: Subsequent antibody staining was performed by repeating antigen retrieval and subsequent steps, with primary antibodies for Tom1 (1:300, sc-514430, Santa Cruz Biotechnology, USA) and Tnnt2 (1:200, 15513-1-AP, Proteintech, Wuhan, China) applied sequentially.

Techniques: Expressing, Gene Expression, Immunofluorescence, Staining, Control, Labeling, Fluorescence

Journal: Frontiers in Cardiovascular Medicine

Article Title: Herb pair of Astragali Radix-Descurainiae Semen attenuate heart failure through the myosin VI-Tom1 complex mediated autophagy

doi: 10.3389/fcvm.2025.1599746

Figure Lengend Snippet: Mode of action, 3D, active pocket, and 2D maps of 10 compounds in MYO6-Tom1 complex and AR-DS decoction. The compounds were Ononin (A) , Astragaloside-IV (B) , Rutin (C) , Folic-acid (D) , Daidzein (E) , Isorhamnetin (F) , Quercetin (G) , Beta-Sitosterol (H) , Kaempferol (I) , Formononetin (J) .

Article Snippet: Subsequent antibody staining was performed by repeating antigen retrieval and subsequent steps, with primary antibodies for Tom1 (1:300, sc-514430, Santa Cruz Biotechnology, USA) and Tnnt2 (1:200, 15513-1-AP, Proteintech, Wuhan, China) applied sequentially.

Techniques:

Journal: Frontiers in Cardiovascular Medicine

Article Title: Herb pair of Astragali Radix-Descurainiae Semen attenuate heart failure through the myosin VI-Tom1 complex mediated autophagy

doi: 10.3389/fcvm.2025.1599746

Figure Lengend Snippet: AR-DS may regulate myocardial autophagy in HF rats through the MYO6-Tom1 complex. (Created using Figdraw ).

Article Snippet: Subsequent antibody staining was performed by repeating antigen retrieval and subsequent steps, with primary antibodies for Tom1 (1:300, sc-514430, Santa Cruz Biotechnology, USA) and Tnnt2 (1:200, 15513-1-AP, Proteintech, Wuhan, China) applied sequentially.

Techniques:

Journal: BMC Cardiovascular Disorders

Article Title: SIRT5 induces autophagy and alleviates myocardial infarction via desuccinylation of TOM1

doi: 10.1186/s12872-024-04120-6

Figure Lengend Snippet: SIRT5 is downregulated in H/R-indued MIRI cell model. (A) mCherry-GFP-LC3 fluorescence was detected to show the autophagy condition in cardiomyocytes before or after H/R induction. (B) The autophagy-related protein markers were assessed by western blot in cardiomyocytes before or after H/R induction. (C) PI positive cells were detected to observe the cell death before or after H/R induction. D-E. SIRT5 expression changes were evaluated through measuring both mRNA and protein levels with RT-qPCR and western blot. ** P < 0.01: H/R vs. CON

Article Snippet: Short hairpin RNAs (shRNAs) targeting SIRT5 (shSIRT5) or TOM1 (shTOM1) as well as their corresponding negative controls (shNC) were all synthesized by Ribobio (Guangzhou, China).

Techniques: Fluorescence, Western Blot, Expressing, Quantitative RT-PCR

Journal: BMC Cardiovascular Disorders

Article Title: SIRT5 induces autophagy and alleviates myocardial infarction via desuccinylation of TOM1

doi: 10.1186/s12872-024-04120-6

Figure Lengend Snippet: Overexpression of SIRT5 promotes autophagy while inhibits cell death of MIRI cell model. (A) The vector expressing SIRT5 was transfected into model cells and the overexpression efficiency was indicated by RT-qPCR data. (B) The autophagy was detected in H/R induced cardiomyocytes after SIRT5 overexpression through mCherry-GFP-LC3 fluorescence analysis. (C) Cell death in H/R induced cardiomyocytes after SIRT5 overexpression. (D) The autophagy-related protein markers were assessed by western blot in H/R induced cardiomyocytes after SIRT5 overexpression. ** P < 0.01: H/R vs. CON; ## P < 0.01: H/R + SIRT5 vs. H/R + vector

Article Snippet: Short hairpin RNAs (shRNAs) targeting SIRT5 (shSIRT5) or TOM1 (shTOM1) as well as their corresponding negative controls (shNC) were all synthesized by Ribobio (Guangzhou, China).

Techniques: Over Expression, Plasmid Preparation, Expressing, Transfection, Quantitative RT-PCR, Fluorescence, Western Blot

Journal: BMC Cardiovascular Disorders

Article Title: SIRT5 induces autophagy and alleviates myocardial infarction via desuccinylation of TOM1

doi: 10.1186/s12872-024-04120-6

Figure Lengend Snippet: SIRT5 interacts with TOM1 and decreases the succinylation of TOM1 protein at site K48. A. The level of TOM1 and succinylated TOM1 in was separately measured cardiomyocytes with SIRT5 overexpression or knockdown by western blot. B-C. The interaction between SIRT5 and TOM1 was demonstrated by exogenous and endogenous co-IP. D. The potential site of TOM1 succinylation was K48 site. E. The effects of SIRT5 overexpression or knockdown on the level of TOM1 or succinylated TOM1 were detected by western blot after K48 site was mutated (K48R). F. The effects of SIRT5 overexpression or knockdown on the protein stability of TOM1 were detected in CHX-treated cardiomyocytes by western blot. ** P < 0.01: SIRT5 vs. vector; ## P < 0.01: shSIRT5 + vs. shNC

Article Snippet: Short hairpin RNAs (shRNAs) targeting SIRT5 (shSIRT5) or TOM1 (shTOM1) as well as their corresponding negative controls (shNC) were all synthesized by Ribobio (Guangzhou, China).

Techniques: Over Expression, Knockdown, Western Blot, Co-Immunoprecipitation Assay, Plasmid Preparation

Journal: BMC Cardiovascular Disorders

Article Title: SIRT5 induces autophagy and alleviates myocardial infarction via desuccinylation of TOM1

doi: 10.1186/s12872-024-04120-6

Figure Lengend Snippet: Silencing of TOM1 reverses the tendencies of autophagy and apoptosis altered by SIRT5 overexpression in MIRI cell model. (A) TOM1 expression was interfered, as validated by RT-qPCR. ** P < 0.01: shTOM1 + vs. shNC. (B) The effect of TOM1 silencing on SIRT5 overexpression-induced autophagy was determined by analyzing mCherry-GFP-LC3 fluorescence. (C) The cell death prevented by SIRT5 overexpression in H/R-induced MIRI cell model was further augmented by after TOM1 knockdown. (D) The changes in autophagy-related protein markers were observed through western blot analysis. ** P < 0.01: H/R vs. CON. ## P < 0.01: H/R + shSIRT5 + shTOM1 vs. H/R + shSIRT5 + shNC

Article Snippet: Short hairpin RNAs (shRNAs) targeting SIRT5 (shSIRT5) or TOM1 (shTOM1) as well as their corresponding negative controls (shNC) were all synthesized by Ribobio (Guangzhou, China).

Techniques: Over Expression, Expressing, Quantitative RT-PCR, Fluorescence, Knockdown, Western Blot

Journal: BMC Cardiovascular Disorders

Article Title: SIRT5 induces autophagy and alleviates myocardial infarction via desuccinylation of TOM1

doi: 10.1186/s12872-024-04120-6

Figure Lengend Snippet: SIRT5 alleviates myocardial injury in in vivo MI models. (A) TTC staining for infarct area of myocardial tissues in four different rat models. (B) HE staining results for pathology of myocardial tissues in four different rat models. (C) The levels of SIRT5, TOM1, TOM1-succ, and autophagic markers (LC3II/1 and p62) was measured by western blot in myocardial tissues obtained from four groups of rat models

Article Snippet: Short hairpin RNAs (shRNAs) targeting SIRT5 (shSIRT5) or TOM1 (shTOM1) as well as their corresponding negative controls (shNC) were all synthesized by Ribobio (Guangzhou, China).

Techniques: In Vivo, Staining, Western Blot