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. 2017 Mar 20;36(9):1215–1226. doi: 10.15252/embj.201695861

Figure EV3. Adult progenitor cells give rise to both neurons and glia (related to Figs 4 and 5).

Figure EV3

  • A
    miR‐31a‐Gal4 stocks lacking FRT42D were crossed to UAS‐CD8‐GFP,hsFlp;FRT42D, tubGal80 flies. Heat shock was induced 1 day post‐eclosion and the flies dissected 1 day later. The bright green spots seen in the images are not correlated with DAPI, indicating that these are non‐specific bright GFP spots. The control shows that the clones that we observe in the MARCM stocks are not phantom clones as no clones are observed in the no‐FRT42D control.
  • B, C
    G‐Trace [UAS‐RFP, UAS‐Flp, Ubi‐p63E(FRT.Stop)GFP] lineage analysis of miR‐31a‐Gal4‐expressing cells in the adult. Gal4 activity was limited to adult stages with Gal80ts. RFP represents ongoing Gal4 activity (or recent activity allowing for perdurance). GFP is a permanent lineage tag for cells that have expressed Gal4. (B) Anti‐elav was used to label neurons. Yellow arrowheads point to RFP+ elav+ cells. White arrowheads point to GFP+ elav+ cells. (C) Anti‐repo was used to label glia. White arrowheads point to GFP+ repo+ cells.
  • D
    Insc‐Gal4‐expressing cells visualized with UAS‐Histone‐RFP gave rise to elav‐expressing neurons (purple; white arrowheads) and repo‐expressing glia (green; yellow arrowheads). Elav+ RFP+ and repo+ RFP+ cells represent non‐overlapping populations.
  • E
    Images showing the central brain regions of flies carrying G‐Trace [UAS‐RFP, UAS‐Flp, Ubi‐p63E(FRT.Stop)GFP] with Insc‐Gal4 and Gal80 ts. Flies were reared at 18°C until eclosion. Flies that were not shifted to the permissive temperature of 29°C (left) showed very few GFP‐expressing clones in the central brain. Flies that were reared right after eclosion for 7 days at 29°C had more clones than those that had been reared for only 1 day at 29°C.
  • F–H
    Flies carrying G‐Trace with the Insc‐Gal4 driver and Gal80 ts were raised at 18°C until the indicated age, after which Gal4 was activated by shifting to 29°C for a further 7 days. Brains were labelled with anti‐repo or anti‐elav. RFP expression represents ongoing or recent Gal4 activity. GFP is a permanent lineage tag for cells that have expressed Gal4. Ctrl indicates the Canton S control background. KO indicates the miR‐31a mutant background. (F) Images of brains shifted to activate Gal4 at 7, 14 and 21 days. Clones expressing repo and GFP are labelled with white arrowheads. (G) Cells expressing elav and RFP represented as a percentage of RFP‐expressing cells. Although it appears that there may be a trend towards fewer elav‐expressing cells in the earlier time points in the mutant brains, the scatter in the data is large, and the differences were not statistically significant. (H) Cells expressing repo and RFP represented as a percentage of RFP‐expressing cells. The number of cells expressing repo was significantly increased in the mutant brains. Unpaired Student's t‐test was used. Error bars represent SEM.