Fig 2: Constitute Lv-shPAK7 silences PAK7 in osteosarcoma cell line. (A) Transfection rate was detected by fluorescence microscope, GFP expression ratio was almost 100%. (B) Validation of PAK7 mRNA level in osteosarcoma cells with RT-PCR analysis; ß-actin was used as an internal control. (C) Validation of PAK7 expression in osteosarcoma cells with Western blot analysis, GAPDH was used as an internal control.

Fig 3: Inhibition of PAK5 remarkably suppresses cell viability in glioma cells with high PAK5 expression. (A-C) Cell viability curves of U87 (A), SHG-44 (B) and CHG-5 (C) cells during the 5 days were evaluated by the MTT assay. The data represent the mean ± SEM of three independent experiments. *, P < 0.05 vs. cells infected with Scr-shRNA lentivirus. (D) PAK5 protein expression in U87 cells infected with a vector with expressing PAK5 after PAK5-shRNA1-mediated PAK5 knockdown. (E) Cell viability curves of U87 cells of three groups during the 5 days were evaluated by the MTT assay. The data represent the mean ± SEM of three independent experiments. *, P < 0.05 vs. cells infected with Scr-shRNA lentivirus.

Fig 4: Overexpression of PAK5 in OS lung metastasis tissue. (A) Reverse transcription-quantitative polymerase chain reaction analysis of the PAK5 mRNA transcript levels in 65 primary OS tissue samples and 13 lung metastasis tissues. (B) PAK5 expression levels detected using immunohistochemistry. Upper row, ×200 magnification. Lower row, ×400 magnification. (C) Kaplan-Meier estimator analysis of survival of 65 patients with OS comparing high and low PAK5 expression (P=0.036). PAK5, p21-activated kinase 5; OS, osteosarcoma.

Fig 5: miR-489 expression is negatively correlated with PAK5 expression and predicts a more favorable clinical outcome in patients with glioma. (A) Relative RNA expression levels of miR-489 in 40 glioma and adjacent matched healthy tissues. miR-489 expression was lower in glioma tissues compared with the matched adjacent tissues. **P<0.01 vs. adjacent tissues. (B) Kaplan-Meier survival analysis demonstrated that upregulated miR-489 expression levels were associated with longer overall survival in patients with glioma. (C) mRNA expression levels of PAK5 in the 40 glioma specimens and adjacent tissues. PAK5 expression was higher in the glioma tissues compared with that in matched adjacent tissues. **P<0.01 vs. adjacent tissues. (D) Kaplan-Meier survival analysis revealed that upregulated expression of PAK5 was associated with shorter overall survival in patients with glioma. (E) Association between relative RNA expression levels of miR-489 and PAK5 in 40 patients. PAK5, p21-activated kinase 5.