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 mitigates hypoxia-mediated oxidative stress in placenta. (A) MDA content in placenta. (B) SOD activity in placenta. (C) Nrf2 mRNA expression levels in placenta. (D) Representative western blotting images. (E) Total Nrf2 expression in placental tissues. (F) Nuclear Nrf2 expression in placental tissues. (G) Nrf2 expression in cytoplasm of placental tissues. (H) Representative immunofluorescence images and (I) quantification of Nrf2 in placenta. Scale bar, 50 μ m. (J) Relative mRNA expression levels of HO-1 and NQO-1. Data are expressed as the mean ± SD. Normal, n=6; IUGR, n=6; IUGR + MOTS-c, n=5. * P<0.05 vs. normal, ** 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; SOD, superoxide dismutase; MDA, malondialdehyde; Nrf2, nuclear factor erythroid 2-related factor 2; HO-1, heme oxygenase 1; NQO-1, NAD(P)H quinone dehydrogenase 1.

Article Snippet: For Nrf2 immunofluorescence staining in HUVECs, cells were fixed with 100% methanol for 15 min at −20°C, followed by permeabilization with 0.1% Triton X-100 for 10 min and blocked with 1% bovine serum albumin (BSA) (cat. no. HY-D0842; MedChemExpress) for 60 min at room temperature.

Techniques: Activity Assay, Expressing, Western Blot, Immunofluorescence

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 attenuates oxidative stress in HUVECs. (A) Relative intensity of DCFH-DA staining. (B) Cellular SOD activity. (C) Cellular MDA content. (D) Representative images of MitoSOX staining. Scale bar, 10 μ m. (E) Representative images of JC-1 staining. Scale bar, 50 μ m. (F) Representative immunofluorescence images of Nrf2 in HUVECs. Scale bar, 5 μ m. (G) Relative nuclear to cytoplasm fluorescence ratio. Results are representative of three independent experiments. Data are expressed as the mean ± SD, n=4. *** P<0.001, # P<0.05 vs. PBS under hypoxic conditions, ## P<0.01 vs. PBS under hypoxic conditions; ### P<0.001 vs. PBS under hypoxic conditions. SOD, superoxide dismutase; MDA, malondialdehyde; DCFH-DA,2'7'-dichlorodihydro fluorescein diacetate ; Nrf2, nuclear factor erythroid 2-related factor 2.

Article Snippet: For Nrf2 immunofluorescence staining in HUVECs, cells were fixed with 100% methanol for 15 min at −20°C, followed by permeabilization with 0.1% Triton X-100 for 10 min and blocked with 1% bovine serum albumin (BSA) (cat. no. HY-D0842; MedChemExpress) for 60 min at room temperature.

Techniques: Staining, Activity Assay, Immunofluorescence, Fluorescence

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: Nrf2 inhibitor ML385 offset the protective effect of MOTS-c against hypoxia-induced dysregulated angiogenesis and oxidative stress in HUVECs. (A) Representative images and (B) quantitative analysis of the in vitro tube formation. Scale bar, 10 μ m. (C) Representative images of DCFH-DA staining and (D) quantification of cellular ROS. Scale bar, 50 μ m. Results are representative of three independent experiments. Data are expressed as the mean ± SD. * P<0.05, ** P<0.01. NS, not significant; DCFH-DA,2',7'-dichlorodihydro fluorescein diacetate; Nrf2, nuclear factor erythroid 2-related factor 2.

Article Snippet: For Nrf2 immunofluorescence staining in HUVECs, cells were fixed with 100% methanol for 15 min at −20°C, followed by permeabilization with 0.1% Triton X-100 for 10 min and blocked with 1% bovine serum albumin (BSA) (cat. no. HY-D0842; MedChemExpress) for 60 min at room temperature.

Techniques: In Vitro, Staining

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: Nrf2 overexpression does not enhance the beneficial effects of MOTS-c on hypoxia-stimulated HUVECs. (A) Relative Nrf2 mRNA expression levels. (B) Cell viability. (C). Representative images of DCFH-DA staining and (D) quantification of cellular ROS. Scale bar, 10 μ m. Data are expressed as the mean ± SD. * P<0.05, ** P<0.01, *** P<0.001, # P<0.05, ## P<0.01. NS, not significant; Nrf2, nuclear factor erythroid 2-related factor 2; ROS, reactive oxygen species; OE, overexpression; DCFH-DA, 2',7'-dichlorodihydro fluorescein diacetate.

Article Snippet: For Nrf2 immunofluorescence staining in HUVECs, cells were fixed with 100% methanol for 15 min at −20°C, followed by permeabilization with 0.1% Triton X-100 for 10 min and blocked with 1% bovine serum albumin (BSA) (cat. no. HY-D0842; MedChemExpress) for 60 min at room temperature.

Techniques: Over Expression, Expressing, Staining

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: For Nrf2 immunofluorescence staining in HUVECs, cells were fixed with 100% methanol for 15 min at −20°C, followed by permeabilization with 0.1% Triton X-100 for 10 min and blocked with 1% bovine serum albumin (BSA) (cat. no. HY-D0842; MedChemExpress) for 60 min at room temperature.

Techniques: Staining, Western Blot, Expressing

Journal: Materials Today Bio

Article Title: 3D printing combined with thermally induced phase separation for engineering hierarchical osteogenic PLA scaffolds

doi: 10.1016/j.mtbio.2025.102621

Figure Lengend Snippet: Macf1 promotes osteogenesis by modulating the cytoskeleton. (A) SEM images of WT and Macf1-KD BMSCs seeded on PLA-196 scaffolds, scale bar = 10 μm. (B) Elongation rate and spreading area of WT and Macf1-OE BMSCs. (C) SEM images of WT and Macf1-OE BMSCs seeded on PLA4 scaffolds, scale bar = 10 μm. (D) Elongation rate and spreading area of WT and Macf1-OE BMSCs. (E) Expression levels of post-Macf1-OE osteogenic marker proteins on PLA4 scaffolds. (F) Expression levels of osteogenic marker proteins following MACF1 on PLA-196 scaffolds. ns: p > 0.05,*: p < 0.05.

Article Snippet: Multiple sections were prepared from the center of the bone defect area, and the sections were subjected to routine hematoxylin and eosin (H&E) staining, as well as immunohistochemical staining for Opn (1:200, Proteintech, China), Ocn (1:200, Proteintech, China), and immunofluorescence staining for Macf1 (1:500, Proteintech, China), Col-I (1:500, Proteintech, China), Cd31 (1:500, Proteintech, China), and CD86 (1:200, Cell Signaling Technology, USA).

Techniques: Expressing, Marker

Journal: Materials Today Bio

Article Title: 3D printing combined with thermally induced phase separation for engineering hierarchical osteogenic PLA scaffolds

doi: 10.1016/j.mtbio.2025.102621

Figure Lengend Snippet: Spatiotemporal expression profiles of osteogenic markers in SD rat calvarial defect models. (A) Immunofluorescence staining of Macf1 at 2 weeks post-implantation, scale bar = 200 μm. (B) Quantification of Macf1 relative fluorescence intensity. (C) Immunofluorescence staining of Col-I at 4 and 8 weeks, scale bar = 200 μm. (D) Quantification of Col-1 relative fluorescence intensity in. (E) Immunofluorescence staining of CD31 expression (neovascular marker) at 4 and 8 weeks, scale bar = 200 μm. (F) Quantification of CD31 relative fluorescence intensity. ns: p > 0.05, *: p < 0.05.

Article Snippet: Multiple sections were prepared from the center of the bone defect area, and the sections were subjected to routine hematoxylin and eosin (H&E) staining, as well as immunohistochemical staining for Opn (1:200, Proteintech, China), Ocn (1:200, Proteintech, China), and immunofluorescence staining for Macf1 (1:500, Proteintech, China), Col-I (1:500, Proteintech, China), Cd31 (1:500, Proteintech, China), and CD86 (1:200, Cell Signaling Technology, USA).

Techniques: Expressing, Immunofluorescence, Staining, Fluorescence, Marker

Journal: Discover Oncology

Article Title: Lipid metabolism-related genes correlate with immune microenvironment and regulate the efficacy of immunotherapy via ferroptosis in melanoma

doi: 10.1007/s12672-025-04163-x

Figure Lengend Snippet: Overexpression ACSL4 increases the therapeutic efficacy of anti-PD-1 antibody in vivo. (A) A schematic view of the treatment plan, (B) Images of isolated tumors from xenograft mice that received indicated treatment. Tumor volumes and weights in each group were calculated and displayed on the right. (C) Immunofluorescence staining of ferroptosis marker PTGS2 in isolated xenograft tumors with indicated treatment. Scale bar = 100 μm. Data represent the mean ± SEM of triplicates. P value was calculated by two-tailed Student’s t-test. ** P < 0.01, *** P < 0.001. n = 6

Article Snippet: The primary antibodies and dilutions for western blotting (WB) and immunofluorescence (IF) staining analysis are listed below: ACSL4 (22401-1-AP, Proteintech, Wuhan, China; 1:100 for IHC, 1:100 for IF and 1:5 000 for WB), ALOX5 (10021-1-Ig, Proteintech; 1:400 for IHC, and 1:500 for WB), PTGS2 (66351-1-Ig, Proteintech; 1:400 for IF), CD8α(ab22378,, Abcam; 1:300 for IF), ACTIN (66009-1-Ig, Proteintech; 1:10 000 for WB).

Techniques: Over Expression, Drug discovery, In Vivo, Isolation, Immunofluorescence, Staining, Marker, Two Tailed Test