Quantification and specificity of monoclonal anti-Pax6 effects. Eye area (measured in arbitrary units using Lucia G software, and distributed among 12 classes) of embryos injected at the blastula stage and presented as cumulative frequency plots. Reagents were always compared pair-wise on a single clutch. (a) Monoclonal anti-Pax6 (HaP6, number of eyes measured n = 161) versus vehicle (phosphate-buffered saline (PBS), n = 164). (b) HaP6 injected first in one half of the clutch (n = 128) versus monoclonal anti-myc (Hamyc, n = 120) injected immediately after in the second half of the clutch. (c) Hamyc (n = 70) versus HaP6 (n = 98), injected in the reverse order to (b). (d) HaP6 (n = 216) versus HaP6 pre-incubated with the amino-terminal arm of Pax6 (HaP6+Nter, n = 132). (e) HaP6 pre-incubated with the amino-terminal arm of Pax6 (HaP6+Nter, n = 198) versus Hamyc pre-incubated with the amino-terminal arm of Pax6 (Hamyc+Nter, n = 138). Differences are statistically significant for (a-d) (p < 0.0001, Mann-Whitney tests).