Fig. 4. DjCHC-RNAi treatment caused CNS atrophy. (A) DjCHC-RNAi treatment caused atrophy of the head and brain from 7 to 14 days after the injection of intact animals. (B) Numbers of head-atrophied planarians in control, X-ray-irradiated, DjCHC-RNAi-treated and DjCHC-RNAi plus X-ray-irradiated planarians. n=30. (C) BrdU-labeling of the trunk piece of control and DjCHC-RNAi-treated planarians. Samples were fixed 4 or 48 hours after BrdU treatment. a, anterior; p, posterior; ph, pharynx. (D) Frequency of mitosis in uncut planarians detected using anti-H3P. Confocal single projection of the fluorescent image of H3P-positive cells (magenta) and the CNS of the identical animal visualized using anti-DjSYT (green) are shown. Control, DjCHC-RNAi-treated and X-ray-irradiated planarians were fixed 14 days after RNAi treatment or irradiation. Animals are oriented with anterior towards the top. (E) The number of labeled mitoses determined using anti-H3P in planarians. Each column represents the average number (±s.e.m.) of anti-H3P-positive cells from 12 planarians. Similar data were obtained in experiments using BrdU. ^***P<0.01. NS, not significant. Scale bars: 100 µm.