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Fig. 4. Semiquantitative RT-PCR demonstrates late zygotic transcription of Hro-hh. (A) Digital images of ethidium bromide-stained agarose gels. Fragments of Hro-hh and 18S rRNA were amplified in separate reactions carried out in parallel. (B) Amount of Hro-hh mRNA during development, relative to stage 10 (100%). At each stage, the intensity of the Hro-nos band was normalized against the 18S rRNA fragment (see Materials and Methods for details). Each reaction sample contained template cDNA equivalent to 4 embryos at the stage indicated [stage 4b ~10 hours AZD, stage 6a ~19 hours AZD, stage 7 ~40 hours AZD, early/mid stage 8 (E/M8) ~65 hours AZD, late stage 8 (L8) ~88 hours AZD, early stage 9 (E9) ~112 hours AZD, late stage 9 (L9) ~140 hours AZD, stage 10 ~150 hours AZD, late stage 10/stage 11(L10/11) 170~185 AZD, late stage 11(L11) ~195 AZD]. Squares indicate the data obtained from the gels shown in (A); circles and triangles represent data obtained starting with independent sets of embryos; black circles show the data from independent PCR.