Fig. 3.

Localization of the folate transport systems in brain parenchyma: A astrocytes, B neurons, C microglia. Mouse brain sections were stained with the following: DAPI nuclear marker, AE390 anti-RFC (1:50) (Panel 1), anti-PCFT (1:50) (Panel 2), or anti-FRα (1:50) (Panel 3). The following cell markers were used to determine cell-specific localization: anti-GFAP (1:50) (astrocyte marker), anti-NeuN (1:100) (neuronal marker), or anti-IBA1 (1:100) (microglial marker). No primary antibody was used as a negative control (Panel 4). Arrows denote localization of RFC, PCFT, or FRα with cell markers. Sections were visualized using confocal microscopy (LSM 700; Carl Zeiss) operated with ZEN software using an oil-immersion 63× lens