Fig. 5. Molecular analysis of DjCHC-RNAi-treated planarians during regeneration. (A-D) Expression patterns of Djnlg (A), DjvlgA (B), ndk (C) and brain-specific homeobox genes (D) were analyzed by whole-mount in situ hybridization 12, 24 or 48 hours after amputation, as indicated. DjCHC-RNAi-treated planarians showed no differences in the expression patterns of Djnlg, DjvlgA, ndk, DjotxA, DjotxB or Djotp compared to those of control animals. (E) Schematic illustration of the discrete expression pattern of the brain-specific homeobox genes. (F) Defects of DjCHC-RNAi-treated planarians 72 hours after amputation were detected in the brain architecture using anti-DjSYT and anti-DjGß. Arrowheads, inverted U-shaped structure; arrows, lateral branches in regenerated blastema. The dashed line indicates the border of the newly formed region and old stump region. (G) Whole-mount immunostaining with VC-1 anti-planarian visual-neuron monoclonal antibody 7 days after regeneration. Scale bars: 50 µm.