Figure 2.

Decreased expression of leukodystrophy-associated Pol III subunits in OPCs increases the proportion of proliferating cells but does not affect migration. (A) Schematic of the transwell insert migration assay. (B) Quantification of siRNA-treated OPC migration, as determined by cell count in a randomly imaged 4X field. Data represents mean $\pm$ SEM from three independent experiments (each presented as an average of 3–6 technical replicates). (C) Representative immunofluorescence images (from three independent experiments with similar results) of siRNA-treated OPCs in proliferation media, showing PDGFRα (green), NG2 (red), and Ki67 (yellow). Nuclei stained with DAPI. Scale bar, 50 μm (20X). (D) Quantification of the % PDGFRα+ (left) and NG2+ (right) cells that are Ki67+ from three independent experiments. (E) Representative images (from five independent experiments with similar results) of siRNA-treated OPCs labelled with EdU. Nuclei stained with DAPI. Scale bar, 50 μm (20X). (F) Quantification of EdU+ cells as a percentage of DAPI following siRNA-treatment. Data represents mean $\pm$ SEM from five independent experiments (each presented as an average of 5 technical replicates), *p < 0.05, **p < 0.01, unpaired, two-sided Student’s t-test.