Fig. 5

The presence of 5mC in miRNA-181a-5p abolishes its funtions. a Caspase-3 activity was measured to estimate apoptosis induction. Cells were co-treated with the indicated miRNA and ABT737 (1 μM) or control. Caspase-3 activity was determined using the Caspase 3 Assay Kit (Abcam, France). b Cell invasion determined by Collagen-Based Cell Invasion Assay (Millipore, France). c Cell Proliferation estimated by cell counting (Countess™ Automated Cell Counter (ThermoFisher, France)). d Impact of the methylation of miRNA-181a-5p on its tumor suppressor function and on BIM expression. The diagram illustrates the experimental procedures. Graphs illustrate the results obtained from 5 mice in each experimental condition. Pictures are representative of tumors obtained for each treatment. BIM expression was quantified using ELISA. e Graph illustrates the stratification of GBM patient samples according to their miRNA-181a-5p expression and methylation levels. Blue, open circles correspond to the patients whose miRNA-181a-5p was unmethylated and highly-expressed (UH). Red, open circles represent the patients with low expression of an unmethylated miRNA-181a-5p (UL). Red, closed circles represent the patients with a methylated miRNA-181a-5p (M). f Overall survival rates in the GBM patient subgroups (Kaplan-Meier) according to miRNA-181a-5p expression and the methylation status as described in Fig. 3i. Patients presenting low expression of an unmethylated miRNA-181a-5p (UL) and a methylated miRNA (M) were included in the same subgroups as these two “signatures” had a low effect on BIM