Fig 1: Inhibition of c-Jun blocks the promoting effects of MAPK15 overexpression on the migration and invasion of osteosarcoma cells. Osteosarcoma cells were divided into three groups: NC group, MAPK15 overexpression (MAPK15-U) group and MAPK15 overexpression plus SP600125 (40 µM) treatment group (MAPK-U+SP600125). (A) Wound-healing assay to detect the migration ability of each group in 143B and MG63 cells. Scale bar, 100 µm. (B) Statistical chart showing the migration ability of each group in 143B and MG63 cells. (C) Transwell assay to detect the invasion ability of each group in MG63 and 143B cells. Scale bar, 100 µm. (D) Statistical chart showing the number of invasion cells per field of each group in 143B and MG63 cells. **P<0.01. MAPK15, mitogen-activated protein kinase 15; NC, negative control.
Fig 2: MAPK15 significantly regulates C-Jun/MMPs signaling pathway. (A) Western blotting was used to detect the expression levels of MAPK15, c-Jun, P-c-Jun, MMP2 and MMP9. MAPK15 expression was inhibited. (B) Statistical analysis for MAPK15, c-Jun, p-c-Jun, MMP2 and MMP9. (C) Western blotting was used to detect the expression of MAPK15, c-Jun, P-c-Jun, MMP2 and MMP9. (D) Statistical analysis for MAPK15, c-Jun, p-c-Jun, MMP2 and MMP9. **P<0.01. MAPK15, mitogen-activated protein kinase 15; MMP, matrix metalloproteinase; NC, normal control; p-, phosphorylated-.
Fig 3: MAPK15 was most significantly upregulated in metastatic OS tissues. (A) Venn diagram showing intersected genes of core genes in the gene module associated with the metastasis of OS and DEGs between metastatic and non-metastasis OS tissues. (B) RT-qPCR was used to detect the expression of FOXJ1 in metastatic (n=15) and non-metastasis (n=11) OS tissues. *P<0.05. (C) RT-qPCR was used to detect the expression of DLEC1 in metastatic (n=15) and non-metastasis (n=11) OS tissues. (D) qRT-PCR was used to detect the expression of MAPK15 in metastatic (n=15) and non-metastatic (n=11) OS tissues. **P<0.01. (E) Immunohistochemistry was performed to detect the expression of MAPK15 in metastatic (n=15) and non-metastasis (n=11) OS tissues. Scale bar, 100 µm. MAPK15, mitogen-activated protein kinase 15; DEG, differentially expressed gene; OS, osteosarcoma; RT-qPCR, reverse transcription-quantitative PCR.
Fig 4: Inhibition of MAPK15 decreases MG63 and 143B cell migration and invasion in vitro. (A) Wound-healing assay was used to detect the migration ability of the sh-scramble and sh-MAPK15 groups in 143B and MG63 cells. Sale bar, 100 µm. (B) Statistical chart showing the migration ability of the sh-scramble and sh-MAPK15 groups in 143B and MG63 cells. (C) Transwell assays were used to detect the invasion ability of both groups in MG63 and 143B cells. Scale bar, 100 µm. (D) Statistical chart showing the number of invasive cells per field. **P<0.01. MAPK15, mitogen-activated protein kinase 15; sh-, short hairpin (RNA).
Fig 5: Inhibition of MAPK15 significantly decreases the metastasis of 143B cells tumors in vivo. (A) Rate of mice free from pulmonary embolism mediated breathing and movement disorders before experiment end. (B) Typical examples of the lungs in the sh-scramble and sh-MAPK15 group mice. (C) Statistical chart showing the metastatic lungs of mice in the sh-scramble and sh-MAPK15 groups. (D) Typical images of H&E staining exhibiting the metastasis of 143B cells in lungs of the sh-scramble and sh-MAPK15 group mice. Scale bar, 1,000 µm. **P<0.01. MAPK15, mitogen-activated protein kinase 15; sh-, short hairpin (RNA).
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