Journal: International Dental Journal

Article Title: 4632427E13Rik Facilitates Jaw Marrow-Derived Mesenchymal Stem Cells Osteogenesis and Angiogenesis Under Hypoxia Through miR-34a-5p/Aldoa/Hif-1α Pathway

doi: 10.1016/j.identj.2025.109364

Figure Lengend Snippet: Aldoa promotes osteogenesis and angiogenesis via the ERK/Hif-1α pathway. A, Alizarin red staining (14 days) and ALP staining (7 days). B, The gene expression levels of Runx2, Alp and Ocn were determined by qRT-PCR. β-actin was used as an internal reference gene. C, Tube formation assay was performed in the presence of CM. The scale bars represent 100 μm. D, The mRNA expression levels of Vegf and CD31 were analysed by quantitative RT-PCR. β-actin was used as an internal reference gene. E, Immunofluorescence staining was performed to detect Vegf expression after being treated by CM. The scale bars represent 25 μm. F, A Kyoto Encyclopaedia of Genes and Genomes (KEGG) enrichment analysis was performed to determine the top related pathways involving these differentially expressed mRNAs. G and H, The protein expression levels of Aldoa, Hif-1α and the phosphorylated of ERK, JUK and p38 among hypoxia, normoxia and hypoxia + si-Aldoa groups were determined by western blot. Semiquantitative analysis of the p-ERK/ERK, p-JUK/JUK and p38/p-p38 ratios were shown. β-actin was used as an internal reference gene. All data were expressed as means ± SD. * P < .05, ** P < .01.

Article Snippet: Rabbit anti-Aldoa (1:1000, 11217-1-AP), rabbit anti-Hif-1α (1:1000, ab179483), rabbit anti-Runx2 (1:1000, 8486S), rabbit anti-CD31 (1:1000, 77699S), rabbit anti-Ocn (1:1000, bs-4917R), rabbit anti-Alp (1:1000, bs-1535R), rabbit anti-Vegf (1:1000, bs-1313R), rabbit anti-ERK (1:1000, bsm-33337M), rabbit anti-p-ERK (1:1000, bs-1646R), rabbit anti-p38 (1:1000, bs-0637R), rabbit anti-p-p38 (1:1000, bs-0636R), rabbit anti-JNK (1:1000, bs-2592R), rabbit anti-p-JNK (1:1000, bs-1640R), and mouse anti-β-actin (1:2000, AF7018) were purchased from Proteintech, Abcam, Cell Signalling Technology, and Bioss, respectively.

Techniques: Staining, Gene Expression, Quantitative RT-PCR, Tube Formation Assay, Expressing, Immunofluorescence, Western Blot

Journal: International Dental Journal

Article Title: 4632427E13Rik Facilitates Jaw Marrow-Derived Mesenchymal Stem Cells Osteogenesis and Angiogenesis Under Hypoxia Through miR-34a-5p/Aldoa/Hif-1α Pathway

doi: 10.1016/j.identj.2025.109364

Figure Lengend Snippet: Aldoa promotes osteogenesis and angiogenesis via the ERK/Hif-1α pathway. A, Alizarin red staining (14 days) and ALP staining (7 days). B, The gene expression levels of Runx2, Alp and Ocn were determined by qRT-PCR. β-actin was used as an internal reference gene. C, Tube formation assay was performed in the presence of CM. The scale bars represent 100 μm. D, The mRNA expression levels of Vegf and CD31 were analysed by quantitative RT-PCR. β-actin was used as an internal reference gene. E, Immunofluorescence staining was performed to detect Vegf expression after being treated by CM. The scale bars represent 25 μm. F, A Kyoto Encyclopaedia of Genes and Genomes (KEGG) enrichment analysis was performed to determine the top related pathways involving these differentially expressed mRNAs. G and H, The protein expression levels of Aldoa, Hif-1α and the phosphorylated of ERK, JUK and p38 among hypoxia, normoxia and hypoxia + si-Aldoa groups were determined by western blot. Semiquantitative analysis of the p-ERK/ERK, p-JUK/JUK and p38/p-p38 ratios were shown. β-actin was used as an internal reference gene. All data were expressed as means ± SD. * P < .05, ** P < .01.

Article Snippet: Rabbit anti-Aldoa (1:1000, 11217-1-AP), rabbit anti-Hif-1α (1:1000, ab179483), rabbit anti-Runx2 (1:1000, 8486S), rabbit anti-CD31 (1:1000, 77699S), rabbit anti-Ocn (1:1000, bs-4917R), rabbit anti-Alp (1:1000, bs-1535R), rabbit anti-Vegf (1:1000, bs-1313R), rabbit anti-ERK (1:1000, bsm-33337M), rabbit anti-p-ERK (1:1000, bs-1646R), rabbit anti-p38 (1:1000, bs-0637R), rabbit anti-p-p38 (1:1000, bs-0636R), rabbit anti-JNK (1:1000, bs-2592R), rabbit anti-p-JNK (1:1000, bs-1640R), and mouse anti-β-actin (1:2000, AF7018) were purchased from Proteintech, Abcam, Cell Signalling Technology, and Bioss, respectively.

Techniques: Staining, Gene Expression, Quantitative RT-PCR, Tube Formation Assay, Expressing, Immunofluorescence, Western Blot

Journal: Translational Oncology

Article Title: Zeaxanthin targets TOP2A to regulate autophagy and suppress lung cancer progression via the MAPK/ERK pathway

doi: 10.1016/j.tranon.2025.102658

Figure Lengend Snippet: Zea modulates MAPK/ERK signaling to inhibit LC progression. A: Analysis of gene expression differences pre- and post-Zea treatment; B: Heat map analysis of TOP 50 DEGs; C: GO functional enrichment of differentially expressed genes; D: KEGG pathway enrichment of differentially expressed genes; E: WB analysis of MAPK/ERK pathway proteins. A549 cells: NC, Zea, Zea + Ro 67–7476. F: Cell viability measured by CCK-8; G: Cell proliferation assessed by colony formation; H: Cell migration analyzed by Transwell; I: Cell apoptosis detected by flow cytometry; J: WB of autophagy-related proteins; K: IF analysis of autophagosomes and lysosomes, with LC3 for autophagosomes and LAMP1 for lysosomes. ** P < 0.01, *** P < 0.001. The data is presented as mean ± standard deviation, and inter-group comparisons are conducted using ANOVA and Tukey’s post hoc test.

Article Snippet: We also wondered whether the impact of Zea on LC cells was associated with the MAPK/ERK signaling, so we treated A549 cells with Zea and subsequently added MAPK/ERK pathway activator Ro 67–7476 (MCE, USA) to activate this pathway.

Techniques: Gene Expression, Functional Assay, CCK-8 Assay, Migration, Flow Cytometry, Standard Deviation

Journal: Translational Oncology

Article Title: Zeaxanthin targets TOP2A to regulate autophagy and suppress lung cancer progression via the MAPK/ERK pathway

doi: 10.1016/j.tranon.2025.102658

Figure Lengend Snippet: Zea targets TOP2A to enhance autophagy via the MAPK/ERK pathway and impedes LC progression. A549 cells: oe-NC, oe-TOP2A, A: TOP2A mRNA level detected by qRT-PCR A549 cells: oe-NC, oe-TOP2A, oe-NC + Zea, oe-TOP2A + Zea. B: WB analysis of TOP2A, MAPK/ERK pathway proteins (p-ERK1/2, ERK1/2, p-MEK, MEK), and autophagy-related proteins (LC3-I, LC3-II, Beclin 1); C: IF detection of autophagosome and lysosome formation; D: CCK-8 assay of cell viability; E: Colony formation assay of cell proliferation; F: Transwell assay of cell migration; G: Flow cytometry analysis of cell apoptosis. ** P < 0.01, *** P < 0.001. The data is presented as mean ± standard deviation, and inter-group comparisons are conducted using ANOVA and Tukey’s post hoc test.

Article Snippet: We also wondered whether the impact of Zea on LC cells was associated with the MAPK/ERK signaling, so we treated A549 cells with Zea and subsequently added MAPK/ERK pathway activator Ro 67–7476 (MCE, USA) to activate this pathway.

Techniques: Quantitative RT-PCR, CCK-8 Assay, Colony Assay, Transwell Assay, Migration, Flow Cytometry, Standard Deviation

Journal: Translational Oncology

Article Title: Zeaxanthin targets TOP2A to regulate autophagy and suppress lung cancer progression via the MAPK/ERK pathway

doi: 10.1016/j.tranon.2025.102658

Figure Lengend Snippet: In vivo evidence that Zea targets TOP2A to influence the MAPK/ERK pathway and autophagy to mitigate LC progression. Animal groups: oe-NC, oe-TOP2A, oe-NC + Zea, oe-TOP2A + Zea. A: Tumor images from the mouse model; B: Weight of mouse tumor tissues; C: Volume of mouse tumor tissues; D: HE staining images of mouse tumor tissues; E: IHC detection of TOP2A and KI67 expression levels in tumor tissues; F: WB analysis of MAPK/ERK pathway proteins (p-ERK1/2, ERK1/2, p-MEK, MEK) and autophagy-related proteins (LC3-I, LC3-II, Beclin 1); G: IF detection of autophagosome and lysosome colocalization. *** P < 0.001. The data is presented as mean ± standard deviation, and inter-group comparisons are conducted using ANOVA and Tukey’s post hoc test.

Article Snippet: We also wondered whether the impact of Zea on LC cells was associated with the MAPK/ERK signaling, so we treated A549 cells with Zea and subsequently added MAPK/ERK pathway activator Ro 67–7476 (MCE, USA) to activate this pathway.

Techniques: In Vivo, Staining, Expressing, Standard Deviation