Figure 8. Nxt1 is required for full expression of tMAC dependent reporter constructs.

(A) Schematic diagram to illustrate reporter construct design. Promoter regions are narrow coloured boxes, transcribed regions are broad boxes. The djl promoter and UTRs are light green; ORF grey; intron dark green. CG42355 promoter and UTRs are pale pink; ORF grey; introns dark pink. UAS-djl-LacZ uses 5×UAS (red) and hsp70 minimal promoter to the TSS (orange) as the promoter. All reporter transcripts comprise 5′ UTR (from djl or CG42355) fused to Adh 5′UTR (cyan) LacZ ORF (blue) and SV40 3′UTR (cyan). (B) RNA in situ hybridisation with a LacZ probe reveals reduced expression of reporter constructs in Nxt1^z2-0488/Nxt1^DG05102 mutant testes compared to control. (C) q-RT-PCR to test LacZ reporter expression in mutant spermatocytes compared to controls. Expression for each transgene was normalised as 1 in the control, and the relative expression in Nxt1^z2-0488 homozygotes (blue) and comr (red) mutants was calculated. (D) q-RT-PCR to test the effect of inclusion of introns on reporter expression in Nxt1^z2-0488 homozygote spermatocytes compared to controls. Expression for each transgene was normalised to 1 in the control samples and relative expression in mutant was calculated. Introduction of two introns into the reporters increased expression in the mutant testes compared to the no intron version. Two separate reporter lines for transgenes are shown.