Fig S5

Efficient heat-shock overexpression of sema4ab does not alter regenerative neurogenesis in lesioned wild-type larvae.
(A) A map of the heat-shock vector injected to over-express sema4ab and a reporter (mCherry) is shown. (B) Heat-shock leads to detectability of mCherry protein mainly in muscle cells surrounding the injury site (position of ventral spinal cord indicated by mnx1:GFP transgene) at 24 hpl. (C, D) Experimental timeline to assess sema4ab over-expression by qRT-PCR after a single heat shock is shown in (C). qRT-PCR analysis shows that a single heat-shock leads to strongly increased sema4ab expression for at least 16 hours post heat-shock (hpHS, D). (E) Over-expression of sema4ab does not elicit any changes in the number of newly generated neurons after spinal lesion (F; gControl: 8.8 cells per larva ± 0.98; hsp70:sema4ab: 6.4 cells per larva ± 0.86; Mann–Whitney U test: p = 0.3850). Each dot for qRT-PCR represents a pool of 50 larvae. β-actin was used as housekeeping gene. Raw data for qRT-PCR can be found in S5 and S6 Data. Error bars show SEM. Dotted lines show the injury site in B and E. Scale bars: 50 µm. Data files for graphs available in S3 Data.