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