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: Research

Article Title: Lymph Node Metastasis-Associated Spatiotemporal Mapping of the TFF3-Linked Niche in Breast Cancer: Integrating Radiogenomic Signatures with Immune-Ecosystem Remodeling

doi: 10.34133/research.1016

Figure Lengend Snippet: 6-MP inhibited the proliferation and LNM of breast cancer (BRCA) cells in vitro. (A) The eXtreme Sum (XSum) algorithm identified potential small molecules and drugs that could correct biological effects caused by dysregulated TFF3 gene expression in BRCA, based on data from the Connectivity Map‌ (cMAP) database. Each scatter point represents a distinct compound, with the y axis showing similarity scores for 1,288 compounds, derived by comparing gene-related features using the XSum method. Compounds with lower scores may inhibit gene-mediated oncogenic effects. (B) qRT-PCR was performed to assess the messenger RNA (mRNA) expression levels of TFF3 after treatment with 5 μM/10 μM 6-MP. (C to E) CCK-8, colony formation, and EdU assays demonstrated the proliferative capacity of MDA-MB-468 cells following treatment with 5 μM/10 μM 6-MP and TFF3 knockdown. (F) Transwell assays assessed the mobility of MDA-MB-468 cells following treatment with 5 μM/10 μM 6-MP and TFF3 knockdown (scale = 200 μm). (G) HLEC tube formation assays demonstrated the lymphangiogenic function of MDA-MB-468 cells following treatment with 5 μM/10 μM 6-MP and TFF3 knockdown (scale = 200 μm). (H to K) The bar charts illustrate the differences observed in colony formation (H), EdU (I), Transwell assays (J), and HLEC tube formation assays (K). (L and M) The levels of EMT proteins were examined following treatment with 5 μM/10 μM 6-MP and TFF3 knockdown by Western blot (L). p-ERK1/2, p-p38, p-JNK, and total ERK1/2, p38, and JNK were analyzed by Western blot (M). (N and O) Rescue experiment validating the functional specificity of 6-MP targeting TFF3 . Migration assays demonstrated that 6-MP inhibited the migration of breast cancer cells, while TFF3 overexpression partially reversed this inhibitory effect (N). The reversal of 6-MP’s action by TFF3 overexpression supports the functional specificity of the 6-MP– TFF3 interaction (O). * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: The antibodies used were rabbit anti-glyceraldehyde-3-phosphate dehydrogenase (1:5,000, Proteintech, China), rabbit anti-TFF3 (1:500, HUABIO, China), rabbit anti-MMP2 (1:1,000, Proteintech, China), rabbit anti-N-cadherin (1:1,000, Zenbio, China), rabbit anti-ERK1/2 (1:2,000, Proteintech, China), rabbit anti-p-ERK1/2 (1:1,000, Proteintech, China), rabbit anti-p38 (1:1,000, Proteintech, China), rabbit anti-p-p38 (1:1,000, Proteintech, China), mouse anti-JNK (1:2,000, Proteintech, China), and rabbit anti-p-JNK1/2/3 (1:500, Cohesion, China).

Techniques: In Vitro, Gene Expression, Derivative Assay, Quantitative RT-PCR, Expressing, CCK-8 Assay, Knockdown, Western Blot, Functional Assay, Migration, Over Expression