Journal: International Journal of Molecular Medicine

Article Title: MOTS-c protects against placental injury via Nrf2 activation in hypoxia-induced intrauterine growth restriction mice

doi: 10.3892/ijmm.2025.5697

Figure Lengend Snippet: MOTS-c administration protects against hypoxia-induced fetal growth restriction. (A) Schematic timeline of the experimental setup. (B) Morphology of fetal mice on GD17.5. (C) Mean fetal weights within each litter. (D) Placental efficiency, which represents the ratio of fetal to placenta weight. (E) Representative images of H&E staining of placental tissues. Scale bar, 20 μ m. (F) Quantification of placental blood sinus area. (G) Representative IHC images CD31. Scale bar, 50 μ m. (H) Quantification of CD31 positive area. (I) Western blotting analysis of CD31, VEGFA and VEGFR2 protein expression in placenta. Data are expressed as the mean ± SD. Normal, n=6; IUGR, n=6; IUGR + MOTS-c, n=5. ** P<0.01 vs. normal, *** P<0.001 vs. normal; # P<0.05 vs. IUGR; ## P<0.01 vs. IUGR; ### P<0.001 vs. IUGR. IUGR, intrauterine growth restriction; GD, gestational day; VEGFA, vascular endothelial growth factor A; VEGFR2, VEGF receptor 2.

Article Snippet: Following a 2 h blocking step at room temperature with 5% BSA (cat. no. HY-D0842; MedChemExpress) in Tris-buffered saline, the membranes were incubated overnight at 4°C with primary antibodies targeting CD31 (1:1,000; cat. no. 77699s; Cell Signaling Technology, Inc.), vascular endothelial growth factor receptor 2 (VEGFR2; 1:1,000; cat. no. 26415-1-AP; Proteintech), vascular endothelial growth factor A (VEGFA; 1:1,000; cat. no. 66828-1-Ig; Proteintech), Nrf2 (1:1,000; cat. no. 62352; Abcam) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH; 1:5,000; cat. no. 263962; Abcam).

Techniques: Staining, Western Blot, Expressing

Journal: International Journal of Molecular Medicine

Article Title: MOTS-c protects against placental injury via Nrf2 activation in hypoxia-induced intrauterine growth restriction mice

doi: 10.3892/ijmm.2025.5697

Figure Lengend Snippet: MOTS-c exposure promotes angiogenesis in HUVECs. (A) MOTS-c content in HUVECs. (B) Cell morphology under white light. Scale bar, 200 μ m. (C) Representative images and (D) quantitative analysis of the in vitro tube formation. Scale bar, 10 μ m. (E) Relative mRNA expression levels of CD31 , VEGFA and VEGFR2 in HUVECs. Results are representative of three independent experiments. Data are expressed as mean ± SD, n=4. * P<0.05, ** P<0.01, *** P<0.001; # P<0.05 vs. PBS under hypoxic conditions, ## P<0.01 vs. PBS under hypoxic conditions. VEGFA, vascular endothelial growth factor A; VEGFR2, VEGF receptor 2.

Article Snippet: Following a 2 h blocking step at room temperature with 5% BSA (cat. no. HY-D0842; MedChemExpress) in Tris-buffered saline, the membranes were incubated overnight at 4°C with primary antibodies targeting CD31 (1:1,000; cat. no. 77699s; Cell Signaling Technology, Inc.), vascular endothelial growth factor receptor 2 (VEGFR2; 1:1,000; cat. no. 26415-1-AP; Proteintech), vascular endothelial growth factor A (VEGFA; 1:1,000; cat. no. 66828-1-Ig; Proteintech), Nrf2 (1:1,000; cat. no. 62352; Abcam) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH; 1:5,000; cat. no. 263962; Abcam).

Techniques: In Vitro, Expressing

Journal: International Journal of Molecular Medicine

Article Title: MOTS-c protects against placental injury via Nrf2 activation in hypoxia-induced intrauterine growth restriction mice

doi: 10.3892/ijmm.2025.5697

Figure Lengend Snippet: MOTS-c protects against hypoxia-induced placental insufficiency in an Nrf2-dependent manner. (A) Morphology of fetal mice on GD17.5 in WT and Nrf2 KO mice. (B) Placental efficiency, which represents the ratio of fetal to placenta weight. (C) Representative images of H&E staining of placental tissues. Scale bar, 100 μ m. (D) Quantification of the placental blood sinus area. (E) Western blotting analysis of CD31, VEGFA and VEGFR2 protein expression levels in placenta. (F) Relative mRNA expression levels of Pgf , Igf2 , Glut1 , Fatp4 and Snat2 in placenta. Data are expressed as mean ± SD. n=4-6. * P<0.05 vs. normal, ** P<0.01 vs. normal, *** P<0.001 vs. normal, # P<0.05 vs. IUGR, ### P<0.001 vs. IUGR.. NS, not significant; IUGR, intrauterine growth restriction; GD, gestational day; Pgf, placental growth factor; Nrf2, nuclear factor erythroid 2-related factor 2; Igf2, insulin-like growth factor 2; Glut1, glucose transporter type 1; Fatp4, fatty acid transporter 4; Snat2, sodium-dependent neutral amino acid transporter-2; VEGFA, vascular endothelial growth factor A; VEGFR2, VEGF receptor 2.

Article Snippet: Following a 2 h blocking step at room temperature with 5% BSA (cat. no. HY-D0842; MedChemExpress) in Tris-buffered saline, the membranes were incubated overnight at 4°C with primary antibodies targeting CD31 (1:1,000; cat. no. 77699s; Cell Signaling Technology, Inc.), vascular endothelial growth factor receptor 2 (VEGFR2; 1:1,000; cat. no. 26415-1-AP; Proteintech), vascular endothelial growth factor A (VEGFA; 1:1,000; cat. no. 66828-1-Ig; Proteintech), Nrf2 (1:1,000; cat. no. 62352; Abcam) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH; 1:5,000; cat. no. 263962; Abcam).

Techniques: Staining, Western Blot, Expressing

Journal: Materials Today Bio

Article Title: Mechanical energy-charged navigating pump repair natural vascular scaffolds to reverse osteoporosis

doi: 10.1016/j.mtbio.2025.102431

Figure Lengend Snippet: Characterization of the mechanical energy-charged navigating pump (MLE/DMV). A) Schematic diagram of the construction of MLE/DMV. B) The size and TEM images of MVs, DMVs, ML-Exo and MLE/DMV (Scale = 100 μm). C, D) Schematic illustration of the physical mixture and co-incubate of DSPE-PEG 2000 -(DSS) 6 and MVs, and fluorescence microscopy showing colocalization. Red, Dil-labeled MVs; green, FITC-labeled DSPE-PEG 2000 -(DSS) 6 (Scale = 5 μm). E) The zeta potential of MVs, DMVs, ML-Exo and MLE/DMV. F, G) Schematic illustration of the physical mixture and co-extrusion of DMVs and ML-Exo, and fluorescence microscopy showing colocalization. Red, Dil-labeled DMVs; green, PHK67-labeled ML-Exo (Scale = 5 μm). H) SDS-PAGE showing the protein profile of ①marker, ②ML-Exo, ③MLE/DMV, ④DMVs. I) Western blot analysis was performed to examine the expression of exosomal proteins (TSG101, CD81, and CD9) and endothelial cell membrane proteins (VEGFR2 and CXCR4) in ML-Exo, MLE/DMV, and DMVs. J) Size and PDI of MLE/DMV in PBS containing 50 % FBS for 7 days. An individual sample was represented by a dot plot, and data are presented as means ± standard deviation (SD); n = 3. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Anti-TSG101 antibody, anti-CD81 antibody, anti-CD9 antibody, anti-VEGFR2 antibody and anti-CXCR4 antibody were purchased from proteintech (Wuhan, China).

Techniques: Fluorescence, Microscopy, Labeling, Zeta Potential Analyzer, SDS Page, Western Blot, Expressing, Membrane, Standard Deviation

Journal: Journal of Translational Medicine

Article Title: ELF3-regulated PES1 targets VEGFR2 to mediate angiogenesis and retinal inner barrier injury in diabetic retinopathy

doi: 10.1186/s12967-025-07334-0

Figure Lengend Snippet: Integrated RNA-seq and scRNA-seq data analysis to screen key transcription factors. ( A ) Volcano plots showing differentially expressed genes (DEGs) between diabetic retinopathy (DR) and control groups in datasets GSE102485 and GSE160306 . ( B ) Scale-free topology model fit for selecting the optimal soft-thresholding power in WGCNA analysis of GSE102485 . Top: The optimal soft threshold (power = 11) is determined. Bottom: Mean connectivity under different soft thresholds. ( C ) Clustering dendrogram of the top 60% variance genes in GSE102485 . Top: Hierarchical clustering tree. Bottom: Module assignment. ( D ) Module-trait relationships between gene modules and DR/control groups in GSE102485 . Module “MEsalmon” exhibits the highest correlation with DR. ( E ) Scale-free topology model fit for selecting the optimal soft-thresholding power in WGCNA analysis of GSE160306 . Top: Optimal soft threshold (power = 6). Bottom: Mean connectivity. ( F ) Clustering dendrogram of the top 60% variance genes in GSE160306 . Top: Hierarchical clustering tree. Bottom: Module assignment. ( G ) Module-trait relationships between gene modules and DR/control groups in GSE160306 . Module “MEred” shows the highest correlation with DR. ( H ) Upset diagram depicting the intersection of WGCNA module genes, upregulated DEGs in DR, and transcription factors (TFs), yielding six common genes (TFDEGs). ( I ) Uniform manifold approximation and projection (UMAP) visualization of 13 cell types identified by integrating single-cell datasets GSE165784 , GSE245561 , and GSE135922 . ( J ) Heatmap displaying marker genes for each cell type. ( K ) FeaturePlot showing the expression patterns of TFDEGs across cell types. Only PES1 is highly expressed in endothelial cells. ( L ) Violin plots comparing the expression levels of PES1 and VEGFR2 in endothelial cells versus other cell types, highlighting their significant upregulation in endothelial cells

Article Snippet: Primary or secondary antibodies were summarized as follows: PES1(1:5000, 13553-1-AP; Proteintech, Wuhan, China; 1:100, sc-166300; Santa Cruz); VEGFA (1:1000, 19003-1-AP; Proteintech); NOX4 (1:5000, 14347-1-AP; Proteintech); VE-cadherin (1:1000, 66804-1-AP; Proteintech; 1:2000, YM8373; Immunoway, China); Occludin (1:5000, 27260-1-AP; Proteintech); SOD1 (1:10000, 10269-1-AP; Proteintech); VEGFR2 (1:1000, 26415-1-AP; Proteintech; 1:1000, R380984; Zenbio, China); ELF3 (1:1000, R389278; Zenbio); SOX4 (1:1000, CSB-PA022431LA01HU; CUSABIO, China); β-actin (1:3000, GB15003-100; Servicebio, China).

Techniques: RNA Sequencing, Control, Marker, Expressing

Journal: Journal of Translational Medicine

Article Title: ELF3-regulated PES1 targets VEGFR2 to mediate angiogenesis and retinal inner barrier injury in diabetic retinopathy

doi: 10.1186/s12967-025-07334-0

Figure Lengend Snippet: PES1 was significantly activated in endothelial cells and retinas under diabetic conditions. ( A ) UMAP plot demonstrating significant differences in human retinal endothelial cells between DR and normal retinal tissues. ( B , C ) Expression profiles of six key transcription factors in human retinal endothelial cells ( B ), with significant upregulation of PES1 and VEGFR2 in DR patient-derived retinal endothelial cells ( C ). ( D, E ) Expression of six key transcription factors in mouse retinal endothelial cells ( D ), showing significant elevation of Pes1 and Vegfr2 in DR mouse retinal endothelial cells ( E ). ( F ) KEGG pathway enrichment analysis of highly expressed genes in diabetic retinopathy (DR) across 13 cell types. ( G ) KEGG pathway enrichment analysis of highly expressed genes in retinal endothelial cells of patients with DR. ( H ) KEGG pathway enrichment analysis of highly expressed genes in retinal endothelial cells of DR mice. ( I - L ) The expression of PES1, VEGFA, and VEGFR2 in rat retinas or HRMECs. Student t-test. n = 3/group. * p < 0.05 versus Ctrl or NG. ( M - P ) The expression of PES1, VEGFA, and VEGFR2 in rat retinas or HRMECs. One-way ANOVA. n = 3/group. * p < 0.05 versus DR + NC shRNA or HG + NC siRNA

Article Snippet: Primary or secondary antibodies were summarized as follows: PES1(1:5000, 13553-1-AP; Proteintech, Wuhan, China; 1:100, sc-166300; Santa Cruz); VEGFA (1:1000, 19003-1-AP; Proteintech); NOX4 (1:5000, 14347-1-AP; Proteintech); VE-cadherin (1:1000, 66804-1-AP; Proteintech; 1:2000, YM8373; Immunoway, China); Occludin (1:5000, 27260-1-AP; Proteintech); SOD1 (1:10000, 10269-1-AP; Proteintech); VEGFR2 (1:1000, 26415-1-AP; Proteintech; 1:1000, R380984; Zenbio, China); ELF3 (1:1000, R389278; Zenbio); SOX4 (1:1000, CSB-PA022431LA01HU; CUSABIO, China); β-actin (1:3000, GB15003-100; Servicebio, China).

Techniques: Expressing, Derivative Assay, shRNA

Journal: Journal of Translational Medicine

Article Title: ELF3-regulated PES1 targets VEGFR2 to mediate angiogenesis and retinal inner barrier injury in diabetic retinopathy

doi: 10.1186/s12967-025-07334-0

Figure Lengend Snippet: VEGFR2 is a potential target gene regulated by PES1. ( A , B ) Gene ontology (GO) and KEGG enrichment analysis based on ChIP-seq for differentially expressed genes in HRMECs. ( C ) Gene tracks based on ChIP-seq of PES1 using Integrative Genomics Viewer (IGV) in different groups. ( D ) The enrichment ability of PES1, H3K4me1, and H3K27ac in VEGFR2 enhancer regions. Student t-test. n = 3/group. * p < 0.05 versus IgG. ( E ) The luciferase activity of VEGFR2 enhancer transfected with PES1 overexpress plasmids or control plasmids. Student t-test. n = 3/group. * p < 0.05 versus PES1 NC group. ( F ) The mRNA level of PES1 and VEGFR2 in HRMECs. One-way ANOVA. n = 3/group. * p < 0.05 versus HG + NC siRNA

Article Snippet: Primary or secondary antibodies were summarized as follows: PES1(1:5000, 13553-1-AP; Proteintech, Wuhan, China; 1:100, sc-166300; Santa Cruz); VEGFA (1:1000, 19003-1-AP; Proteintech); NOX4 (1:5000, 14347-1-AP; Proteintech); VE-cadherin (1:1000, 66804-1-AP; Proteintech; 1:2000, YM8373; Immunoway, China); Occludin (1:5000, 27260-1-AP; Proteintech); SOD1 (1:10000, 10269-1-AP; Proteintech); VEGFR2 (1:1000, 26415-1-AP; Proteintech; 1:1000, R380984; Zenbio, China); ELF3 (1:1000, R389278; Zenbio); SOX4 (1:1000, CSB-PA022431LA01HU; CUSABIO, China); β-actin (1:3000, GB15003-100; Servicebio, China).

Techniques: ChIP-sequencing, Luciferase, Activity Assay, Transfection, Control

Journal: Journal of Translational Medicine

Article Title: ELF3-regulated PES1 targets VEGFR2 to mediate angiogenesis and retinal inner barrier injury in diabetic retinopathy

doi: 10.1186/s12967-025-07334-0

Figure Lengend Snippet: Schematic diagram of the regulatory mechanism of ELF3/PES1/VEGFR2 signaling pathway in DR retinal angiogenesis. ELF3 and PES1 are highly activated in DR. ELF3 promotes PES1 transcription. PES1 enhances VEGFR2 transcription leading to increased angiogenesis. In contrast, PES1 reduces the expression of VE-cadherin and Occludin, promotes vascular endothelial cell dysfunction, disrupts the blood-retinal barrier (BRB), and contributes to the development of DR

Article Snippet: Primary or secondary antibodies were summarized as follows: PES1(1:5000, 13553-1-AP; Proteintech, Wuhan, China; 1:100, sc-166300; Santa Cruz); VEGFA (1:1000, 19003-1-AP; Proteintech); NOX4 (1:5000, 14347-1-AP; Proteintech); VE-cadherin (1:1000, 66804-1-AP; Proteintech; 1:2000, YM8373; Immunoway, China); Occludin (1:5000, 27260-1-AP; Proteintech); SOD1 (1:10000, 10269-1-AP; Proteintech); VEGFR2 (1:1000, 26415-1-AP; Proteintech; 1:1000, R380984; Zenbio, China); ELF3 (1:1000, R389278; Zenbio); SOX4 (1:1000, CSB-PA022431LA01HU; CUSABIO, China); β-actin (1:3000, GB15003-100; Servicebio, China).

Techniques: Expressing