Journal: Journal of Hard Tissue Biology

Article Title: Effect of circular RNA hsa_circ_0008016 on the Osteogenic and Odontogenic Ability of Stem Cells from Apical Papilla

doi: 10.2485/jhtb.33.105

Figure Lengend Snippet: Figure 5. Effect of hsa-circ-0008016 on OS/OD protein expression of SCAPs. A. Protein expression of BSP, DMP1, DSPP, OCN, RUNX2 in dif- ferent groups. B. Grayscale analysis of A. The expressions of OCN and BSP in the experimental group were significantly increased, while DSPP was significantly decreased. The expressions of RUNX2 and DMP1 were not statistically significant. *P < 0.05, **P < 0.01

Article Snippet: Primary antibodies included BSP, DSPP, DMP1, OCN, RUNX2 (Santa Cruz Biotechnology, USA), and beta-actin (Proteintech Group).

Techniques: Expressing

Journal: Journal of Hard Tissue Biology

Article Title: Effect of circular RNA hsa_circ_0008016 on the Osteogenic and Odontogenic Ability of Stem Cells from Apical Papilla

doi: 10.2485/jhtb.33.105

Figure Lengend Snippet: Figure 4. Effect of hsa-circ-0008016 on OS/OD gene expression of SCAPs. A. The transfection efficacy of pLenti-CMV-Has_circ_0008016- GFP-Puro. The qRT-PCR results showed that the relative expression of hsa_circ_0008016 in cells transfected with overexpression vector was significantly higher than that in the control group. B. qRT-PCR showed that the expressions of DMP1 and BSP were significantly increased in the experimental group, while the expressions of COL1 and DSPP were significantly decreased. The expressions of FGFR1, RUNX2, OCN and ALP were not statistically significant. *P < 0.05, **P < 0.01, and ***P < 0.001

Article Snippet: Primary antibodies included BSP, DSPP, DMP1, OCN, RUNX2 (Santa Cruz Biotechnology, USA), and beta-actin (Proteintech Group).

Techniques: Gene Expression, Transfection, Quantitative RT-PCR, Expressing, Over Expression, Plasmid Preparation, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: METTL14 promotes migration and invasion of choroidal melanoma by targeting RUNX2 mRNA via m6A modification

doi: 10.1111/jcmm.17577

Figure Lengend Snippet: RUNX2 is the key target of METTL14 in CM. (A) Correlation of METTL14 and RUNX2 expression in the TCGA database. (B) qRT‐PCR analysis of RUNX2 mRNA after METTL14 inhibition or overexpression. (C) WB results indicate that protein expression of RUNX2 is significantly downregulated or upregulated after METTL14 knockdown or overexpression, respectively.

Article Snippet: The slides were incubated overnight at 4 °C with the primary antibodies anti‐METTL14 (1:1000, HPA038002, Sigma‐Aldrich) and anti‐RUNX2 (1:1000, MAB2006, RD), followed by addition of secondary antibodies to an avidin‐biotin‐peroxidase complex (e.g. biotinylated anti‐rabbit and anti‐mouse antibodies in goats).

Techniques: Expressing, Quantitative RT-PCR, Inhibition, Over Expression

Journal: Journal of Cellular and Molecular Medicine

Article Title: METTL14 promotes migration and invasion of choroidal melanoma by targeting RUNX2 mRNA via m6A modification

doi: 10.1111/jcmm.17577

Figure Lengend Snippet: RUNX2 mRNA can be methylated by METTL14 and it promotes migration and invasion of CM cells in vitro. (A–E) RUNX2 mRNA can be methylated by METTL14. A, MeRIP‐qPCR analysis was used to determine m6A modification enrichment in RUNX2 mRNA after overexpressing METTL14 in MUM‐2B cells. (B) C, RUNX2 methylation site. (D) Mutations at the RUNX2 methylation site. (E) Luciferase activities in MUM‐2B cells transfected with RUNX2‐WT or RUNX2‐Mut + vector or METTL14. (F–J) RUNX2 promotes migration and invasion of CM cells in vitro. (F) Kaplan–Meier survival analysis of CM tumour samples suggest that high RUNX2 expression levels are related to reduced overall survival (OS). (G) Protein expression of RUNX2 in CM and normal choroidal tissues. (H) IHC staining of RUNX2 in CM and normal choroidal tissues. (I, J) Transwell assays were used to determine the effects of RUNX2 on migration and invasion capability in OCM1 and MUM‐2B cells.

Article Snippet: The slides were incubated overnight at 4 °C with the primary antibodies anti‐METTL14 (1:1000, HPA038002, Sigma‐Aldrich) and anti‐RUNX2 (1:1000, MAB2006, RD), followed by addition of secondary antibodies to an avidin‐biotin‐peroxidase complex (e.g. biotinylated anti‐rabbit and anti‐mouse antibodies in goats).

Techniques: Methylation, Migration, In Vitro, Modification, Luciferase, Transfection, Plasmid Preparation, Expressing, Immunohistochemistry

Journal: Journal of Cellular and Molecular Medicine

Article Title: METTL14 promotes migration and invasion of choroidal melanoma by targeting RUNX2 mRNA via m6A modification

doi: 10.1111/jcmm.17577

Figure Lengend Snippet: Role of METTL14 and RUNX2 in metastasis. (A) Knockdown of RUNX2 inhibits lung metastasis in nude mice models. WB verified the successful construction of knockdown stabilized MUM‐2B cells. WT and RUNX2 knockdown MUM‐2B stable cells were injected via the tail vein. Representative images of metastatic lung tumours. (B, C) Transwell assay demonstrates the effects of METTL14 and RUNX2 on the migration and invasion capability of CM cells. Overexpression of RUNX2 partially reversed the reduction in cell migration and invasion caused by decreased METTL14 expression. All data are presented as mean ± SD from three independent replicates. Student's t ‐test was used to assess inter‐group differences.

Article Snippet: The slides were incubated overnight at 4 °C with the primary antibodies anti‐METTL14 (1:1000, HPA038002, Sigma‐Aldrich) and anti‐RUNX2 (1:1000, MAB2006, RD), followed by addition of secondary antibodies to an avidin‐biotin‐peroxidase complex (e.g. biotinylated anti‐rabbit and anti‐mouse antibodies in goats).

Techniques: Injection, Transwell Assay, Migration, Over Expression, Expressing

Journal: Biomaterials advances

Article Title: In vitro development and optimization of cell-laden injectable bioprinted gelatin methacryloyl (GelMA) microgels mineralized on the nanoscale.

doi: 10.1016/j.bioadv.2024.213805

Figure Lengend Snippet: Fig. 5. Osteocyte morphology and functionality in mineralized and non-mineralized microgels. A-B and E-F represent the confocal images of non-mineralized (A-B) and mineralized (E-F) samples. Microgels stained with actin (green), DAPI (blue), and SOST (red) and SOST intensity (normalized fluorescence per cell) in mineralized microgels (A-H, K-L). Also, the dendrite length in μm (I) and the number of dendrites per cell (I-J) were not significantly different between mineralized and non-mineralized samples. Statistical differences are represented by **** p < 0.0001 after the One-way ANOVA test post-Turkey’s corrections.

Article Snippet: Cells were permeabilized with 0.1 % Triton X100, blocked with 1.5 % bovine serum albumin in PBS, and incubated at room temperature for 1 h. Samples were then washed in PBS and incubated with the primary antibody (rabbit M.G. da Costa Sousa et al. Biomaterials Advances 159 (2024) 213805 polyclonal SOST antibody (Novus Biologicals, NBP1-77461)) (1:100 dilution), overnight at 4 ◦C.

Techniques: Staining, Fluorescence