Figure 5.

RT-PCR- and western blot-analysis of vari-mutant alleles. (A) RT-PCR on total RNA of wild-type and different vari-mutant embryos (> 8h), using vari-long- and vari-short-specific primer pairs vari-long-5/3 or vari-short-5/3 (see Fig. 1A). No vari-long transcripts can be detected in embryos homozygous for the deficiency Df(2L)DS6 and the allele vari^F033. vari-long transcripts are slightly more abundant in vari^MD109 and strongly enriched in vari^03953b embryos. Both vari-long and vari-short transcripts are truncated in vari^38EFa2 (black arrowheads). vari-short transcripts are absent in vari^MD109, strongly reduced in vari^F033 and vari^03953b, but only mildly affected in vari^R3, vari³²⁷ and vari^R979. Transcripts from the rp49 gene serve as internal loading control. (B) Western blot analysis of the same mutant alleles as in A. Vari-long protein is present in wild-type amounts in vari^MD109, absent in embryos homozygous for the deficiency Df(2L)DS6 and the allele vari^F033, reduced in vari^03953b,vari³²⁷ and vari^R979 and truncated in vari^R3 (white arrow) and vari^38EFa2 (asterisk). Vari-short protein is absent in vari^MD109, in the deficiency Df(2L)DS6 and in vari^R3 and reduced in the remaining alleles. RT-PCR and western-blot analysis of the deficiency Df(2L)DS6, which removes the vari locus, serves as negative control.