Figure 3
Design and validation of different combinations of miRNAs and their chemosensitizing effects on GBM cells in combination with TMZ or DOX. (a) miRNA loaded PLGA-PEG nanoparticles were synthesized using W/O/W double emulsion method optimized in our lab51. The miRNAs were then tested in different GBM cell lines. MicroRNA combinations were tested using two different assays: PI staining based FACS to estimate apoptotic population and cell cycle analysis, MTT assay to find the cell proliferation and viability, and western blotting assay to find the expression of selected target genes. (b) PI staining based FACS analysis results showing the increase in apoptotic populations after treatment by different combinations of miRNAs as compared to control cells, and cells treated with NPs alone. Numbers shown in red fonts in each subplot are the apoptotic (Apo) populations of the cells as a percentage of total number of cells. (c) MTT assay results of cells pretreated with miRNAs and treated with different concentrations of TMZ. (d) MTT assay results of cells pretreated with miRNAs and treated with different concentrations of DOX. The table shows the presence or absence of each miRNA in different treatment conditions. Results are expressed as a percentage of survival compared to cells treated with control NPs. All the experiments were carried out on U87-MG cells (repeated three times by independent experiments with triplicates in each experiment). Corresponding timeline for each experimental study is shown below as a schematic workflow. Error bars show standard deviations. Significance levels are shown in the right margins where, *represent p-value < 0.05, **represent p-value < 0.01, ***represent p-value < 0.001, ****represent p-value < 0.0001. Asterisks show significant difference compared to control NPs of the corresponding dose.
