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. 2007 Dec 6;104(50):19879–19884. doi: 10.1073/pnas.0708806104

Fig. 2.

Fig. 2.

Early exposure to Shh is critical to increase chondrogenesis by CNCCs in mass cultures. (A) The mean number (± SEM) of cartilage nodules per d6 culture of mes-rhombencephalic NCCs is shown in the absence and presence of 100 ng/ml Shh added during the first 48 h (d0–d2) or from d2 to d6 (n = 28; **, P < 0.001). (B) Chondrogenesis by CNCCs isolated after 15- or 24-h migration from explanted mesencephalon, and anterior rhombencephalon (R1–R2) is given by mean number (± SEM) of cartilage nodules per d6 culture grown with and without 100 ng/ml d0–d2 Shh addition (n = 10; **, P < 0.001; *, P < 0.05). (C) Quantification of Sox9+ cells (mean number ± SEM) in d2 cultures of mesencephalic NCCs (harvested after 15 h) grown in control medium alone (−Shh) or with addition of either 100 ng/ml Shh (+Shh) or 5 μM cyclopamine (−Shh + Cyc) or both (+Shh + Cyc) (n = 4; **, P < 0.001). As determined by counting Hoechst-stained quail nuclei, the mean total number of cells did not significantly differ between culture conditions (data not shown).