Fig. 4

Injury-induced Müller glia divide asymmetrically once to generate neurogenic clusters. (A) Sequential labeling with EdU and BrdU. Two predicted alternative results are depicted: if Müller glia (MG) divide only once, they incorporate only EdU (green); if they undergo multiple divisions, they incorporate both EdU and BrdU (green+magenta=white). (B) Some nGFP⁺ (green) nuclei in flat-mounted retinas at 42 hpl are EdU⁺ (red). (C) Pair of EdU⁺ (red) nuclei at 42 hpl, of which only one is PCNA⁺ (magenta). (D) Optical z-stack projection at 72 hpl: clusters of BrdU⁺ nuclei (magenta) with few EdU⁺ nuclei (green). Right panels: single optical section of EdU⁺/nGFP⁺/BrdU^- nucleus (arrows). (E) Lateral view of BrdU⁺ nuclei (magenta) clusters with a single EdU⁺ (green)/BrdU-nucleus, reconstructed from optical Z-stack. Right panel: BrdU⁺ clusters (magenta outline). (F) Cluster with single EdU⁺ cell (white/red) at 72 hpl. (G) Cluster of weakly EdU⁺ cells (white/red) with one strongly EdU⁺ cell. (H) Cluster with one PCNA^-/strongly GFP⁺ (green) cell (arrows) and PCNA⁺ (magenta)/weakly GFP⁺ progenitors. Scale bars: 10 μm.