Figure 1—figure supplement 3.

Figure 1—figure supplement 3. fluorescence activated cell sorting (FACS) and gating strategies.

(A) Flow cytometry plots of dissociated, uninjured retinae from Tg(pcna:EGFP);Tg(gfap:mCherry) fish and gating strategy for sorting calcein-positive cells. Plot 1 displays all the events in a single cell suspension according to the forward (x axis) and side scatter (y axis) areas. Among cells gated in 1, singlets (P2) were chosen by considering the area (x axis) and the height (y axis) of the forward scatter in 2. Height (x axis) and width (y axis) of the side scatter were plotted in 3 to further gate singlets among P2. Finally, only calcein-positive (i.e. living) cells were gated among singlets in 4. The gating strategy was the same for the uninjured as well as all lesioned samples. Samples consisted of single cell suspensions derived from 8 retinae of 4 fish (2 males and 2 females) each. (B) Flow cytometry plots showing EGFP and mCherry fluorescence of cells from WT, Tg(gfap:mCherry) and Tg(pcna:EGFP) dissociated retinae.