Figure 2.

Expression of MANF in the retina. A, Expression of MANF during postnatal development. Retinas were collected at designated time points from wild-type Sprague Dawley rats. Levels of MANF were detected by Western blot analysis with anti-MANF antibodies. High levels of MANF expression were detected during postnatal development (from PD1 to PD16; A, upper panel). As the retinas mature, the expression decreases (from PD25 to PD60; A, upper panel). The same blot was reprobed with anti-β-actin antibodies as loading controls (A, lower panel). Cryosections of 12 µm of eyes of PD20 wild-type Sprague Dawley rats along the vertical meridian were stained with anti-MANF antibodies (B, D, E). Müller cells were identified by antibodies against GS (C–E). Extensive colocalization of MANF and GS immunoreactivities (D, E, yellow) indicates that MANF in the retina is expressed in Müller cells. RPE cells, neurons in the GCL, and neurons in the INL also display MANF immunoreactivities (B, D, white arrowheads in E). OS, outer segments; IS, inner segments; OPL, outer plexiform layer; IPL, inner plexiform layer. Scale bar: 50 µm (D) and 20 µm (E).