Journal: Frontiers in Cell and Developmental Biology

Article Title: Dialogue between nutrition and reproduction: preliminary exploration of sperm quality response to high-fat diet in mice

doi: 10.3389/fcell.2025.1640714

Figure Lengend Snippet: Western blot analysis and quantification of apoptosis, autophagy, and oxidative stress markers. Proteins were extracted from sperm samples of the control diet (CD, n = 3) and high-fat diet (HFD, n = 3) groups using RIPA buffer. Equal amounts of protein (20 μg) were separated via 12% SDS–PAGE and transferred to PVDF membranes (350 mA, 90 min). Membranes were blocked with 5% BSA and incubated overnight at 4 °C with primary antibodies against Bax, Bcl-2, Beclin-1, Caspase-8, Caspase-9, LC3A/B, Mn SOD (all 1:1,000, CST), and β-Actin (1:2,000). After incubation with HRP-conjugated secondary antibodies (1:5,000) and ECL detection, protein bands were visualized and quantified using ImageJ. Densitometric values were normalized to β-Actin. Quantitative results are shown as bar graphs for each marker, illustrating the differential expression between groups. Note: *: P < 0.05, **: P < 0.01, ***: P < 0.001 vs. CD.

Article Snippet: Equal amounts (20 μg) of protein were boiled in 4× SDS sample buffer at 100 °C for 10 min, separated by 12% SDS-PAGE, and transferred to PVDF membranes (Millipore, IPVH00010) at 350 mA for 90 min. Membranes were blocked with 5% BSA in TBST (0.1% Tween-20) for 1 h at room temperature and then incubated overnight at 4 °C with the following primary antibodies:Bax (1:1,000, Cell Signaling Technology, CST#2772), Bcl-2 (1:1,000, CST#3498), Caspase-3 (1:1,000, CST#9662), Caspase-8 (1:1,000, CST#9746), Caspase-9 (1:1,000, CST#9508), Mn SOD (1:1,000, Abcam, ab13533), Beclin-1 (1:1,000, CST#3495), LC3B (1:1,000, CST#3868), β-Actin (1:2,000, CST#3700) as the internal control.

Techniques: Western Blot, Control, SDS Page, Incubation, Marker, Quantitative Proteomics

Journal: Scientific Reports

Article Title: CXCL8 mediates macrophage migration in canine oral malignant melanoma

doi: 10.1038/s41598-025-22749-x

Figure Lengend Snippet: mRNA expression levels of chemokine in seven canine melanoma cell lines. a CCL5 , b CX3CL1 , c CSF-1 , d IL-34 , ( e ) CXCL8 , and ( f ) CCL2 . The relative expression level was normalized to the RPL32 gene . g Correlation between the number of migratory RAW264.7 and CXCL8 mRNA expression levels. h Production of CXCL8 in seven canine melanoma cell lines. i Correlation between the number of migratory RAW264.7 and CXCL8 secretion. All data are presented as means ± SD of three independent experiments.

Article Snippet: The recombinant canine CXCL8 protein (NBP2-34,908, Novus Biologicals, Centennial, CO, USA) was supplemented at a final concentration of 200 ng/mL into the CM.

Techniques: Expressing

Journal: Scientific Reports

Article Title: CXCL8 mediates macrophage migration in canine oral malignant melanoma

doi: 10.1038/s41598-025-22749-x

Figure Lengend Snippet: a , b Relative number of RAW264.7 cells migrated by CMeC1 wild-type CM or CMeC1 CXCL8 knockout CM. c Relative number of RAW264.7 cells migrated by CMeC1 CM. The CM of CMeC1 wild-type cells were treated with anti-dog CXCL8 (1 μg/ml) or mouse IgG isotype (1 μg/ml). All data are presented as means ± SD of three independent experiments. ** P < 0.01; *** P < 0.001.

Article Snippet: The recombinant canine CXCL8 protein (NBP2-34,908, Novus Biologicals, Centennial, CO, USA) was supplemented at a final concentration of 200 ng/mL into the CM.

Techniques: Knock-Out

Journal: Scientific Reports

Article Title: CXCL8 mediates macrophage migration in canine oral malignant melanoma

doi: 10.1038/s41598-025-22749-x

Figure Lengend Snippet: Relative number of RAW264.7 cells migrated by CMeC1 CM. The CM of CMeC1 wild-type or CXCL8 knockout cells were treated with dog CXCL8 recombinant protein (200 ng/ml). All data are presented as means ± SD of three independent experiments. ** P < 0.01; *** P < 0.001.

Article Snippet: The recombinant canine CXCL8 protein (NBP2-34,908, Novus Biologicals, Centennial, CO, USA) was supplemented at a final concentration of 200 ng/mL into the CM.

Techniques: Knock-Out, Recombinant

Journal: Scientific Reports

Article Title: CXCL8 mediates macrophage migration in canine oral malignant melanoma

doi: 10.1038/s41598-025-22749-x

Figure Lengend Snippet: Relative number of canine macrophage DH82 cells migrated by CMeC1 CM. The CM of CMeC1 wild-type or CXCL8 knockout were treated with anti-dog CXCL8 (1 μg/ml) or mouse IgG isotype (1 μg/ml). All data are presented as means ± SD of four independent experiments. ** P < 0.01; *** P < 0.001.

Article Snippet: The recombinant canine CXCL8 protein (NBP2-34,908, Novus Biologicals, Centennial, CO, USA) was supplemented at a final concentration of 200 ng/mL into the CM.

Techniques: Knock-Out