Journal: Current Issues in Molecular Biology

Article Title: Identification and Validation of MTFP1 as a Mitochondrial Target Restoring Dynamics and ECM Remodeling in Acute Myocardial Infarction

doi: 10.3390/cimb48030293

Figure Lengend Snippet: Flowchart of analysis. LASSO, Least Absolute Shrinkage and Selection Operator; ROC, receiver operating characteristics; KEGG, Kyoto Encyclopedia of Genes and Genomes; GO, Gene Ontology; ANN, artificial neural network; GSEA, Gene Set Enrichment Analysis; IHC, immunohistochemistry; RT-QPCR, quantitative real-time polymerase chain reaction; AUC, the area below the curve; MTFP1, mitochondrial fission process 1; DNAJC28, DnaJ heat shock protein family (Hsp40) member C2.

Article Snippet: Following a 30 min room temperature block with 3% bovine serum albumin (BSA), the sections were incubated overnight at 4 °C with primary antibodies, including MTFP1 antibody (proteintech, Cat#:14257-1-AP; dilution ratio 1:200).

Techniques: Selection, Immunohistochemistry, Quantitative RT-PCR, Real-time Polymerase Chain Reaction

Journal: Current Issues in Molecular Biology

Article Title: Identification and Validation of MTFP1 as a Mitochondrial Target Restoring Dynamics and ECM Remodeling in Acute Myocardial Infarction

doi: 10.3390/cimb48030293

Figure Lengend Snippet: GeneMANIA network analysis and single-gene GSEA enrichment analysis. ( A ) GeneMANIA interaction network of key genes. ( B ) GSEA enrichment of DNAJC28. ( C ) GSEA enrichment of MTFP1. The red area (left region) represents the region where genes positively correlated with the target phenotype are enriched. At the left side of the ranked gene list (the end with smaller rank values), genes highly positively correlated with AMI are concentrated. If the peak of the enrichment score curve for a gene set falls on the left side, it indicates that the gene set is primarily enriched among positively correlated genes, potentially promoting the disease process. The blue area (right region) represents the region where genes negatively correlated with the target phenotype are enriched. At the right side of the ranked gene list (the end with larger rank values), genes highly negatively correlated with AMI are concentrated. If the peak of the enrichment score curve for a gene set falls on the right side, it indicates that the gene set is primarily enriched among negatively correlated genes, potentially inhibiting the disease process.

Article Snippet: Following a 30 min room temperature block with 3% bovine serum albumin (BSA), the sections were incubated overnight at 4 °C with primary antibodies, including MTFP1 antibody (proteintech, Cat#:14257-1-AP; dilution ratio 1:200).

Techniques:

Journal: Current Issues in Molecular Biology

Article Title: Identification and Validation of MTFP1 as a Mitochondrial Target Restoring Dynamics and ECM Remodeling in Acute Myocardial Infarction

doi: 10.3390/cimb48030293

Figure Lengend Snippet: Dual network analyses and five targeted drugs for biomarkers. ( A ) Transcription factor-gene regulatory network, showing predicted upstream TFs targeting MTFP1 and DNAJC28 ; each green node represents a TF, with purple edges denoting regulatory interactions. ( B ) Competing endogenous RNA (ceRNA) regulatory network for MTFP1 and DNAJC28 . Orange nodes represent lncRNAs, green nodes represent miRNAs, and purple edges denote miRNA–mRNA and lncRNA–miRNA interactions. Key interactions include mmu-miR-155-5p and mmu-miR-124-3p co-regulating both target genes. A Vina score lower than −5 kcal/mol indicates favorable binding affinity. Notably, epigallocatechin gallate showed the strongest docking affinity with both targets (−7.1/−7.6 kcal/mol). ( C , D ) Predicted docking conformation of epigallocatechin gallate (EGCG) within the MTFP1 and DNAJC28 binding pocket. Electrostatic surface rendering highlights hydrogen bond interactions and hydrophobic contacts.

Article Snippet: Following a 30 min room temperature block with 3% bovine serum albumin (BSA), the sections were incubated overnight at 4 °C with primary antibodies, including MTFP1 antibody (proteintech, Cat#:14257-1-AP; dilution ratio 1:200).

Techniques: Binding Assay

Journal: Current Issues in Molecular Biology

Article Title: Identification and Validation of MTFP1 as a Mitochondrial Target Restoring Dynamics and ECM Remodeling in Acute Myocardial Infarction

doi: 10.3390/cimb48030293

Figure Lengend Snippet: Expression profiles of biomarkers MTFP1 and DNAJC28 in vivo and in vitro models. ( A ) mRNA expression levels of MTFP1 in different cardiac regions of C57/BL6 male mice randomly assigned to sham or MI groups (n = 3). ( B ) Temporal mRNA expression patterns of MTFP1 at different post-MI timepoints. ( C ) mRNA expression levels of DNAJC28 in different cardiac regions. ( D ) Temporal mRNA expression patterns of DNAJC28 at different post-MI timepoints. ( E , F ) Protein expression of MTFP1 in sham and MI groups. ( G , H ) MTFP1 protein expression in cardiomyocytes under normoxia vs. hypoxia at indicated durations. ( I ) MTFP1 mRNA expression in cardiomyocytes during hypoxic exposure. ( J ) Representative images of HE staining (scale bar = 200 μm). Masson’s trichrome (scale bar = 200 μm). Picrosirius red staining (scale bar = 200 μm). MTFP1 immunohistochemistry (scale bar = 200 μm) in sham vs. MI hearts. ( K , L ) Quantification of Collagen Content by Masson and Sirius Red Staining expressed as a percentage. ( M ) Mean optical density of MTFP1 in sham operation group and myocardial infarction group. All experiments were conducted in triplicate with three independent biological replicates per group. Data are presented as mean ± SEM. Statistical analysis was performed using unpaired two-tailed Student’s t -test for two-group comparisons and one-way ANOVA followed by Tukey’s post hoc test for multiple-group comparisons. Significance levels were defined as: p < 0.05 (*), p < 0.01 (**), p < 0.001 (***), and p < 0.0001 (****).

Article Snippet: Following a 30 min room temperature block with 3% bovine serum albumin (BSA), the sections were incubated overnight at 4 °C with primary antibodies, including MTFP1 antibody (proteintech, Cat#:14257-1-AP; dilution ratio 1:200).

Techniques: Expressing, In Vivo, In Vitro, Staining, Immunohistochemistry, Two Tailed Test

Journal: Current Issues in Molecular Biology

Article Title: Identification and Validation of MTFP1 as a Mitochondrial Target Restoring Dynamics and ECM Remodeling in Acute Myocardial Infarction

doi: 10.3390/cimb48030293

Figure Lengend Snippet: MTFP1 overexpression ameliorates cardiac function, reduces infarct size, and mitigates post-MI fibrosis. ( A , B ) Validation of MTFP1 overexpression. C57/BL6 male mice were randomly allocated into: control group (n = 3), GFP group (n = 3), MTFP1 overexpression group (n = 3). MTFP1 expression levels were quantified to confirm successful overexpression. ( C – G ) Echocardiographic Assessment of Cardiac Function. Mice were randomized into: sham group (n = 3), MI group (n = 3), MI + GFP group (n = 3), MI + MTFP1 group (n = 3). Parameters evaluated: Ejection fraction (EF%), fractional shortening (FS%), left ventricular end-diastolic diameter (LVEDD; mm), left ventricular end-systolic diameter (LVESD; mm). ( H , I ) Infarct size quantification by TTC staining. Experimental groups: Sham (n = 3), MI (n = 3), MI + MTFP1 (n = 3). ( J – L ) Histopathological evaluation. Groups analyzed: Sham (n = 3), MI (n = 3), MI + GFP (n = 3), MI + MTFP1 (n = 3). Staining performed: HE staining (scale bar = 200 μm), Masson’s trichrome (scale bar = 200 μm), Picrosirius red (scale bar = 200 μm). All experiments were independently repeated three times. All experiments were conducted in triplicate with three independent biological replicates per group. Data are presented as mean ± SEM. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test for multiple-group comparisons. Significance levels were defined as: p < 0.05 (*), p < 0.01 (**), p < 0.001 (***), and p < 0.0001 (****).

Article Snippet: Following a 30 min room temperature block with 3% bovine serum albumin (BSA), the sections were incubated overnight at 4 °C with primary antibodies, including MTFP1 antibody (proteintech, Cat#:14257-1-AP; dilution ratio 1:200).

Techniques: Over Expression, Biomarker Discovery, Control, Expressing, Staining

Journal: Current Issues in Molecular Biology

Article Title: Identification and Validation of MTFP1 as a Mitochondrial Target Restoring Dynamics and ECM Remodeling in Acute Myocardial Infarction

doi: 10.3390/cimb48030293

Figure Lengend Snippet: MTFP1 restores mitochondrial fission–fusion balance and ECM remodeling via p-DRP1/MMP9/TIMP1 axis in hypoxia-induced cardiomyocytes. ( A , B ) Western blot analysis of myocardial tissue from MI and MI + MTFP1 overexpression (OE) groups assessing MTFP1, DRP1, p-DRP1 (Ser616), MMP9, and TIMP1 expression. Quantification of protein expression is shown (n = 3/group). ( C , D ) Gelatin zymography of myocardial tissue showing pro-MMP9 and active MMP9 levels, with corresponding quantification (n = 3/group). ( E , F ) Western blot analysis of fibrosis-associated proteins COL1A1 and α-SMA in myocardial tissue and corresponding quantification (n = 3/group). ( G , H ) ROS levels in hypoxia-induced H9C2 cell detected by DCFH-DA fluorescence staining and quantitative fluorescence intensity analysis (n = 3/group; scale bar = 200 μm). ( I ) Schematic illustration of the proposed mechanism: MTFP1 overexpression restores mitochondrial dynamics and ECM remodeling by regulating the p-DRP1/MMP9/TIMP1 signaling axis, thereby attenuating fibrosis and oxidative stress while improving cardiac function. All experiments were conducted in triplicate with three independent biological replicates per group. Data are presented as mean ± SEM. Statistical analysis was performed using unpaired two-tailed Student’s t -test for two-group comparisons and one-way ANOVA followed by Tukey’s post hoc test for multiple-group comparisons. Significance levels were defined as: p < 0.01 (**), p < 0.001 (***), and p < 0.0001 (****).

Article Snippet: Following a 30 min room temperature block with 3% bovine serum albumin (BSA), the sections were incubated overnight at 4 °C with primary antibodies, including MTFP1 antibody (proteintech, Cat#:14257-1-AP; dilution ratio 1:200).

Techniques: Western Blot, Over Expression, Expressing, Zymography, Fluorescence, Staining, Two Tailed Test

Journal: ACS Omega

Article Title: Study on Antitumor Secondary Metabolites from Euphorbia ebracteolata Hayata: Structural Characterization, Network Pharmacology, Molecular Docking and In Vitro

doi: 10.1021/acsomega.5c04875

Figure Lengend Snippet: Effect of compound (±)- 1 on apoptosis and cell-cycle profiling in SMMC-7721 cells. Cells were exposed to varying concentrations of compound (±)- 1 (0, 0.1, 0.3, 0.6 μM) for 24 h. (A) Apoptosis of SMMC-7721 cells was assayed by flow cytometry. (B) Cell-cycle profiling was performed using a FACSCanto flow cytometer. Data and profiles for the G0/G1 and G2/M phases, analyzed with Diva software and ModFit LT ver. 3.0, are shown in the gray area. Cells in the S phase are represented as the shaded area. (C) The proportion of apoptotic cells was quantified and expressed as the mean ± SD of three independent experiments (*** p < 0.001). (D) Representative FACS histograms from three independent experiments.

Article Snippet: The Cell Cycle and Apoptosis Analysis Kit, Annexin V-FITC Apoptosis Detection Kit, Mitochondrial Membrane Potential and Apoptosis Detection Kit with Mito-Tracker Red CMXRos and Annexin V-FITC were obtained from Beyotime (Shanghai, China).

Techniques: Flow Cytometry, Software

Journal: Neural regeneration research

Article Title: CHCHD2 Thr61Ile mutation impairs F1F0-ATPase assembly in in vitro and in vivo models of Parkinson's disease.

doi: 10.4103/1673-5374.378010

Figure Lengend Snippet: Figure 2 ｜Cellular ATP levels and F1F0-ATPase activity in isolated mitochondria in a in SH-SY5Y cell model of Parkinson’s disease. (A, B) Flag-vector: LV-Flag-vector (Ubi-MCS-3FLAG-SV40-puromycin vector)-transfected SH-SY5Y cells cultured in MEM-F12 medium; Flag-vector + MPP+: LV-Flag-vector- transfected SH-SY5Y cells treated with MPP+ (500 μM for 24 hours); CHCHD2-Flag + MPP+: LV-CHCHD2-Flag-transfected SH-SY5Y cells treated with MPP+ (500 μM for 24 hours); CHCHD2-T61I + MPP+: LV-CHCHD2-T61I-Flag-transfected SH-SY5Y cells treated with MPP+ (500 μM for 24 hours). (A) Detection of ATP levels in control or MPP+-treated SH-SY5Y cells stably transfected with empty vector or constructs encoding WT or T61I-mutant CHCHD2. (B) Effect of WT or T61I-mutant CHCHD2 on ATP synthase specific activity in control or MPP+-treated cells. (C) Effect of WT or T61I-mutant CHCHD2 on ATP synthase specific activity in AVV-transfected mice with or without MPTP treatment. CHCHD2-Flag + MPTP: AVV-CHCHD2-Flag–transfected mice treated with MPTP (25 mg/kg, twice a week, for 5 weeks); CHCHD2-T61I + MPTP: AVV-CHCHD2-T61I–transfected mice treated with MPTP (25 mg/kg, twice a week, for 5 weeks); Flag-vector: AVV-Flag (CMV- betaGlobin-MCS-3Flag-SV40 polyA (GV411) vector)-transfected mice treated with normal saline (twice a week, for 5 weeks); Flag-vector + MPTP: AVV-Flag-transfected mice treated with MPTP (25 mg/kg, twice a week, for 5 weeks). Data are expressed as the mean ± SD (n = 3 independent experiments). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 (one-way analysis of variance followed by Tukey’s post hoc test). ATP: Adenosine triphosphate; MPP+: 1-methyl-4-phenylpyridinium; MPTP: 1-methyl-4-phenyl-1,2,3,6- tetrahydropyridine.

Article Snippet: Detection of mitochondrial permeability transition pore opening Opening of the mitochondrial permeability transition pore (mPTP) was detected by the calcein-AM/cobalt assay following the manufacturer’s instructions (C2009S, Beyotime Biotechnology).

Techniques: Activity Assay, Isolation, Plasmid Preparation, Transfection, Cell Culture, Control, Stable Transfection, Construct, Mutagenesis, Saline

Journal: Neural regeneration research

Article Title: CHCHD2 Thr61Ile mutation impairs F1F0-ATPase assembly in in vitro and in vivo models of Parkinson's disease.

doi: 10.4103/1673-5374.378010

Figure Lengend Snippet: Figure 3 ｜CHCHD2 T61I mutation promotes mitochondrial permeability transition pore (mPTP) opening in SH-SY5Y cells. CHCHD2-Flag: LV-CHCHD2-Flag-transfected SH-SY5Y cells; CHCHD2-T61I: LV-CHCHD2-T61I-Flag-transfected SH-SY5Y cells; Flag-vector: LV-Flag-vector (Ubi-MCS-3FLAG-SV40-puromycin vector)-transfected SH-SY5Y cells. (A) Calcein-AM and CoCl2 were administered to live cells expressing WT or T61I-mutant CHCHD2, and fluorescence intensity was detected by flow cytometry. (B) Effect of WT or the T61I-mutant CHCHD2 on mPTP opening as determined by detecting mitochondrial membrane potential using JC-1 fluorescent probes (white squares) in control or MPP+-treated SH-SY5Y cells. The fluorescence intensity of intracellular JC-1 aggregates (red) in the MPP+ + CHCHD2-Flag group was greater than that in the MPP+ + Flag-vector group. The CHCHD2-T61I group showed an increased amount of JC-1 monomers (green). Scale bars: 10 μm. (C) The mean fluorescence intensity of JC-1 aggregates or monomers in control or MPP+-treated SH-SY5Y cells expressing WT or T61I-mutant CHCHD2. Data are expressed as the mean ± SD (n = 3 independent experiments). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 (one-way analysis of variance followed by Tukey’s post hoc test). Calcein-AM: Calcein acetoxymethyl ester; MPP+: 1-methyl-4-phenylpyridinium; mPTP: mitochondrial permeability transition pore; WT: wild type.

Article Snippet: Detection of mitochondrial permeability transition pore opening Opening of the mitochondrial permeability transition pore (mPTP) was detected by the calcein-AM/cobalt assay following the manufacturer’s instructions (C2009S, Beyotime Biotechnology).

Techniques: Mutagenesis, Permeability, Transfection, Plasmid Preparation, Expressing, Fluorescence, Flow Cytometry, Membrane, Control

Journal: Neural regeneration research

Article Title: CHCHD2 Thr61Ile mutation impairs F1F0-ATPase assembly in in vitro and in vivo models of Parkinson's disease.

doi: 10.4103/1673-5374.378010

Figure Lengend Snippet: Figure 5 ｜CHCHD2 T61I mutation aggravates movement deficits and nigral DA neuron function in a mouse model of PD. (A) Experimental design. AAV-CHCHD2, AAV-CHCHD2 T61I, or AAV-Vector was stereotaxically injected into the substantia nigra pars compacta, and 4 weeks later mice were injected intraperitoneally with normal saline or MPTP for 5 weeks. One day after the last MPTP/normal saline injection, behavioral tests were performed, and then the mice were sacrificed. Flag-vector: AVV-Flag(CMV-betaGlobin-MCS-3Flag-SV40 polyA (GV411) vector)-transfected mice treated with normal saline (twice a week, for 5 weeks); Flag-vector+MPTP: AVV-Flag– transfected mice treated with MPTP (25 mg/kg, twice a week, for 5 weeks); CHCHD2-Flag+MPTP: AVV-CHCHD2-Flag–infected mice treated with MPTP (25 mg/kg, twice a week, for 5 weeks); CHCHD2-T61I+MPTP: AVV-CHCHD2-T61I–transfected mice treated with MPTP (25 mg/kg, twice a week, for 5 weeks). (B) The grasping test was used to examine grip strength. (C) The pole-climbing test was used to examine bradykinesia. (D) The rotarod test was used to examine motor coordination. (E) Representative immunohistochemical staining of TH-positive neurons in the substantia nigra (SN). There were fewer TH-positive neurons in the SN in the AAV-CHCHD2-T61I + MPTP group than in the AAV-Flag-vector + MPTP and AAV-CHCHD2-Flag + MPTP groups. Scale bars: 800 μm. (F) Quantification of the TH-positive cells shown in E. (G) Representative images of double-immunofluorescent staining for CHCHD2-Flag (green, Alexa Fluor 488) and TH (red, Alexa Fluor 555) (white squares) in the SN. There were significantly fewer TH-positive neurons in the SN in the AAV-CHCHD2-T61I + MPTP group than in the AAV-Flag-vector + MPTP and AAV-CHCHD2-Flag + MPTP groups. Scale bars: 50 μm. (H) Quantification of TH mean fluorescence in the SN as shown in G. (I) Representative immunoblot for TH in the SN. (J) TH expression in the SN. Results are expressed as the mean ± SD (n ≥ 6). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 (one-way analysis of variance followed by Tukey’s post hoc test). ACTB: β-Actin; AAV: adeno-associated virus; DA: dopaminergic; DAPI: 4′,6-diamidino-2-phenylindole; MPTP: 1-methyl-4-phenyl- 1,2,3,6-tetrahydropyridine; TH: tyrosine hydroxylase; SN: substantia nigra.

Article Snippet: Detection of mitochondrial permeability transition pore opening Opening of the mitochondrial permeability transition pore (mPTP) was detected by the calcein-AM/cobalt assay following the manufacturer’s instructions (C2009S, Beyotime Biotechnology).

Techniques: Mutagenesis, Plasmid Preparation, Injection, Saline, Transfection, Infection, Immunohistochemical staining, Staining, Fluorescence, Western Blot, Expressing, Virus

Journal: Neural regeneration research

Article Title: CHCHD2 Thr61Ile mutation impairs F1F0-ATPase assembly in in vitro and in vivo models of Parkinson's disease.

doi: 10.4103/1673-5374.378010

Figure Lengend Snippet: Figure 8 ｜CHCHD2 modulates OSCP expression in MPP+-/MPTP-induced PD models. (A, B) Flag-vector: LV-Flag-vector (Ubi-MCS-3FLAG-SV40-puromycin vector)-transfected SH-SY5Y cells cultured in MEM-F12 medium; Flag-vector+MPP+: LV-Flag-vector–transfected SH- SY5Y cells treated with MPP+ (500 μM for 24 hours); CHCHD2-Flag+MPP+: LV-CHCHD2-Flag–transfected SH-SY5Y cells treated with MPP+ (500 μM for 24 hours); CHCHD2-T61I + MPP+: LV-CHCHD2-T61I-Flag–transfected SH-SY5Y cells treated with MPP+ (500 μM for 24 hours). (A) Representative immunoblot of OSCP, CHCHD2, and ACTB in MPP+-treated SH-SY5Y cells. (B) Quantification of relative OSCP expression shown in A. (C, D) Flag-vector: AVV-Flag(CMV-betaGlobin-MCS-3Flag-SV40 polyA (GV411) vector)-transfected mice treated with normal saline (twice a week, for 5 weeks); Flag-vector + MPTP: AVV-Flag-infected mice treated with MPTP (25 mg/kg, twice a week, for 5 weeks); CHCHD2-Flag + MPTP: AVV-CHCHD2-Flag- infected mice treated with MPTP (25 mg/kg, twice a week, for 5 weeks); CHCHD2-T61I + MPTP: AVV-CHCHD2-T61I–transfected mice treated with MPTP (25 mg/kg, twice a week, for 5 weeks). (C) Representative immunoblot of OSCP, CHCHD2, and ACTB in control or MPTP-treated mice. (D) Quantification of relative OSCP expression shown in C. Data normalized to SH-SY5Y cells overexpressing Flag-vector or C57BL/6J mice transfected with AAV-Flag are expressed as the mean ± SD (n = 3 independent experiments for cells, n = 6 for animals). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 (one-way analysis of variance followed by Tukey’s post hoc test). ACTB: β-Actin; MPP+: 1-methyl-4-phenylpyridinium; MPTP: 1-methyl- 4-phenyl-1,2,3,6-tetrahydropyridine; OSCP: oligomycin sensitivity conferral protein.

Article Snippet: Detection of mitochondrial permeability transition pore opening Opening of the mitochondrial permeability transition pore (mPTP) was detected by the calcein-AM/cobalt assay following the manufacturer’s instructions (C2009S, Beyotime Biotechnology).

Techniques: Expressing, Plasmid Preparation, Transfection, Cell Culture, Western Blot, Saline, Infection, Control