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. 2018 Apr 12;10:775–785. doi: 10.2147/CMAR.S157261

Figure 2.

Figure 2

Overexpression of miR-142-3p increases MGMT promoter methylation in GBM cell lines.

Notes: Methylation-specific PCR assay showing methylation status of MGMT promoter in S1 cells (A) and R1 cells (B) infected with control (shLuc) and miR-142-3p-encoding recombinant lentivirus at low and high multiplicities of infection. Lane U represents amplification of unmethylated MGMT promoter and lane M represents methylated MGMT promoter. Total DNA extracted from PBMCs was used as control: lane U represents amplification of unmethylated PBMC DNA and lane M represents amplification of methylated PBMC DNA pretreated with SssI methyltransferase. qRT-PCR showing overexpression of miR-142-3p in S1 cells (C) and R1 cells (D) after infection with recombinant lentivirus at low (Low) and high (High) multiplicities of infection. Expression levels in miR-142-3p-overexpressing cells quantified relative to expression level in control cells infected with shLuc lentiviral construct, with ΔΔCt quantification with GAPDH amplification used as internal control.

Abbreviations: GBM, glioblastoma multiforme; qRT-PCR quantitative real-time polymerase chain reaction; PBMCs, peripheral blood mononuclear cells.