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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 <t>p38</t> 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.
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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 <t>p38</t> 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.
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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 <t>p38</t> 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.
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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 <t>p38</t> 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.
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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 <t>p38</t> 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.
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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.

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