Figure 7.

Pten acts in RPCs to control responsiveness to amacrine cell negative feedback signals. A, Schematic illustration of aggregation assay protocol. RPCs are shown in green and amacrine cells in red. B–G, The combinations of aggregate conditions are shown in B, D, and F. Immunolabeling of aggregated cell pellets with Pax6 (green) and BrdU (red) is shown with DAPI counterstain (blue). E15.5 RPCs were cultured alone (B, C) or with P2 wild-type (D, E) or Pten cKO (F, G) retinal cells. Arrowheads mark Pax6/BrdU double⁺ nuclei (C, E, G). H, Percentage of BrdU⁺ E15.5 cells differentiated into Pax6⁺ amacrine cells when cultured alone (black bar) or with P2 wild-type (white bar) or Pten cKO (red bar) retinal cells. I–P, Immunolabeling of aggregated cell pellets with Pax6 (green) and BrdU (red) with DAPI counterstain (blue). E15.5 RPCs were cultured alone (I, J) or with P2 wild-type (K, L), and E15.5 Pten cKO RPCs were cultured alone (M, N) or with P2 wild-type (O, P) retinal cells. Arrowheads mark Pax6/BrdU double⁺ nuclei (J, L, N, P). Q, Percentage of BrdU⁺ E15.5 cells differentiated into Pax6⁺ amacrine cells when cultured alone (wild-type, black bar; Pten cKO, white bar) or with P2 wild-type (blue bar, light blue bar) retinal cells. *p < 0.05; **p < 0.01; ***p < 0.001.