Figure 2

Crude HEM enhanced OL maturation. Crude HEM at the indicated concentrations was added to the cultures for 3 days after OPCs had been maintained in DM for 2 days (Fig. 1B). (A) The cultures were subjected to GC and MBP staining (red), followed by DAPI nuclear counterstaining (blue). GC⁺- and MBP⁺-OLs in the cultures were quantified as described in the “Materials and methods” section. Each spot represents as the data quantified from one photo captured from the cultures. Arrows shown in the representative images indicate OLs with the shape of an extended membrane. (B) Total proteins were isolated from cultures treated with crude HEM for 3 days and subjected to Western blot analysis to examine MBP and PLP protein levels. The intensity of the immunoreactive bands shown in the immunoblots was quantified by ImageJ software version 1.52a (https://imagej.nih.gov/ij/) and normalized to the level of GAPDH, which was used as a loading control. The immunoblot images used for quantification are provided in Fig. S5. Data are presented as the means ± SEMs of the three independent experiments. The raw immunoblot images are shown in Fig. S5. *p < 0.05, **p < 0.01 compared with the control culture. Scale bar in (A) 50 μm.