Skip to main content
. 2018 May 4;37(13):e98163. doi: 10.15252/embj.201798163

Figure EV2. Two RNAi lines that target pros with different strengths lead to NSC expansion from early larval stages (related to Fig 2).

Figure EV2

  1. Schematic representation of pros mRNA isoforms: Each box represents an exon; untranslated regions are in gray; and coding regions are in white; in red are the target regions for prosRNAi LH and prosRNAi KK, both of which target all isoforms.
  2. Pictures of WL3 stage DAL lineages (outlined with dashed lines in split magenta channel) in WT or in animals in which Pros has been knocked down with either RNAi at 22°C. Lineages with prosRNAi LH knock‐down contain a few Pros+ cells in contrast to lineages with prosRNAi KK knock‐down, in which no Pros+ cells are detectable and which are larger (single optical sections of stainings performed in parallel and imaged with same settings). In line with the functional data presented in main Fig 2 (panel E), this demonstrates that the two RNA is specifically target pros but that prosRNAi LH is weaker than prosRNAi KK.
  3. Quantification of Dpn+ and Dpn− cells in DAL lineages in the conditions indicated; histograms represent the mean (n = 5–6 CNSs; details in source data) and error bars SEM. Compare with Fig 2B.
  4. Pictures of permanently labeled DAL lineages (outlined in dashed lines in split magenta channel; maximum intensity projections) at adult stage, expressing prosRNAi LH under the strong induction of actin5C‐GAL4. Unlike in animals reared at 22°C, in which EdU was never seen, stronger pros depletion at 25°C results in clones with ectopically replicating cells (boxed area shown at higher magnification in inset). Note that the actin5C promoter responsible for permanent labeling leads to significantly higher levels of expression than the engrailed regulatory sequence. Compare with Fig 2E.
  5. Time course of Dpn+ and Dpn− cells in DAL lineages in the conditions indicated; histograms represent the mean (n = 4–13 CNSs; details in source data) and error bars SEM. Unlike in controls (driver animals reared at 25°C), non‐tumorigenic expansion using prosRNAi LH results in ectopic NSCs already from EL3 stage.
Data information: See main text for abbreviations.Source data are available online for this figure.