Fig. 8

The Transcriptional Activation of YTHDF2 Mediated by PRMT6 Requires Its Methyltransferase Activity; Inhibiting This Activity Suppresses Glioma Cell Migration, Invasion, and EMT. A: Analysis of H3R2me2a, PRMT6, and YTHDF2 protein levels in LN229 and U251 cells treated with varying concentrations of EPZ020411 for 24 h. B: Assessment of YTHDF2 mRNA expression after 24-hour treatment with 20µM EPZ020411 in LN229 and U251 cells. C: Dual-luciferase reporter assays in LN229 or U251 cells treated with 20µM EPZ020411 for 24 h, with or without PRMT6 overexpression. D: Wound healing assay to evaluate cell migration in LN229 and U251 cells with or without EPZ020411 treatment. E-F: Transwell assays assessing cell migration and invasion with or without EPZ020411 treatment. Scale bar = 200 μm. G: Examination of EMT and Wnt-β-catenin pathway-related protein expression in LN229 and U251 cells treated with or without 20µM EPZ020411 for 24 h. H: Wild-type LN229 cells were intracranially implanted into nude mice followed by subcutaneous administration of EPZ or saline for three weeks to observe the effect of EPZ on in vivo tumorigenesis. Representative brain sections stained with H&E displayed xenograft tumors (upper panel, scale bar = 1.5 mm). In vivo invasion assays were conducted by examining the tumor margins in mouse brains (lower panel, scale bar = 100 μm). I: Tumor volumes for each group of mice were calculated. J: The relative invasive fingers of each tumor were calculated under a microscope by counting prominent and diffuse tumor tissues. K: Representative images of immunohistochemical staining for N-cadherin and E-cadherin in xenograft tumors from control and EPZ-treated groups of nude mice. Scale bar = 100 μm