Figure 5

Cell proliferation is required for regeneration of oral structures. (A) Schematic of experiments. (B; E-H; M-P) Nuclei of proliferating cells (green) labeled with the thymidine analog EdU at 96 hpa, and all nuclei counterstained with Hoechst (blue). (C; I-L; Q-T) Nuclei of proliferating cells labeled with EdU at 96 hpa (green). (B, C) Regeneration of oral structures in untreated control polyps 96 hpa. (D) Chart of average tentacle lengths in control and inhibitor treated polyps 96 hpa. Values are the mean of measurements for at least 3 polyps at each time point. Error bars: standard error. Asterisk: p value < 0.05; Student’s t-test of treatment versus control. (E, I) Failure to undergo regeneration of oral structures after treatment with 20 mM hydroxyurea from 18 hpa to 96 hpa. (F, J) Regeneration of oral structures after treatment with 20 mM hydroxyurea from 2 hpa to 18 hpa. (G, K) Regeneration of oral structures after treatment with 20 mM hydroxyurea from 18 hpa to 30 hpa. (H, L) Regeneration of oral structures after treatment with 20 mM hydroxyurea from 30 hpa to 42 hpa. (M, Q) Failure to undergo regeneration of oral structures after treatment with 0.1 μM nocodazole from 18 hpa to 96 hpa. (N, R) Regeneration of oral structures after treatment with 0.1 μM nocodazole from 2 hpa to 18 hpa. (O, S) Failure to undergo regeneration of oral structures after treatment with 0.1 μM nocodazole from 18 hpa to 30 hpa. (P, T) Failure to undergo regeneration of oral structures after treatment with 0.1 μM nocodazole from 30 hpa to 42 hpa. Scale bars = 50 μm.