Figure 6
Addition of CG-8 decreases condensation number and density in vitro and impairs digit development in vivo. (a) Staining by PNA of a 2-day control leg culture (b) Culture treated with 10 μg/ml CG-8, showing a decrease in number of condensations, with a concomitant and uniform decrease in their sizes. (c) Control 2-day culture stained with DAPI. (d) Culture treated with 10 μg/ml rabbit polyclonal anti-CG-8 antibody, fixed at day 2, and stained with DAPI. (e) Graph showing change in condensation number upon addition of 10 μg/ml CG-8 and 10 μg/ml polyclonal anti-CG-8 antibody, respectively. Photomicrographs (a-e) are at same level of magnification; scale bar in (a) represents 0.25 mm. (f) Graph showing change in inter-condensation distance upon addition of 10 μg/ml CG-8 and 10 μg/ml polyclonal anti-CG-8 antibody, respectively. (g) Graph showing change in condensation size both upon addition of CG-8 and polyclonal anti-CG-8 antibody, respectively. The three variables (mean condensation number, mean size and mean inter-condensation distance) were compared as in the legend to Figure 5. (h-i) Wing bud (i) injected with CG-8 on 5th day of incubation, fixed and stained with Alcian blue, reveals absence of any cartilage primordia in the autopod and poorly stained, smaller zeugopodial cartilage in comparison with (h) control left wing. (j-k) Right wing bud injected on 5th day of incubation with (j) CG-8 and (k) contralateral untreated control wing bud, both stained with Alcian blue at day 9, showing decrease in length of zeugopodial primordia due to CG-8 treatment. (l) Graph showing decrease in zeugopodial cartilage primordium length when CG-8 was injected into 5-day wing buds. The primordium lengths were analyzed using one-way analysis of variance comparing means by the Tukey-Kramer post-hoc analysis. Results are shown as mean ± S.E.M. (* p < 0.05).