Figure 4.

Expression patterns of specific PSI-interacting transcripts are affected in v16;P[PSIΔAB] mutants. (A) Smoothened distributions of mean ratios calculated using normalized data from n independent microarray experiments. Distributions for the v16;P[PSIΔAB] versus v16;P[PSI] differential expression experiments (mRNA profiling; adult male RNA samples; n = 9) and for the PSI-interacting experiments (IP αPSI; embryonic RNA samples; n = 5) are both centered and normal-like. (B) Total RNA purified from 0–12-h embryos (∼100 ng, lane 1) or 1/250 of the RNA immunopurified from 1 mL of embryonic nuclear extract using total rabbit IgG (lane 2) or affinity-purified anti-PSI polyclonal antibodies (lane 3; ∼8 ng of RNA) were analyzed by RT–PCR with (+) or without (−) reverse transcription step and PCR primers specific for the indicated cDNA. PCR products were resolved on 1% agarose gels and stained with ethidium bromide. (C) Quantitative RT–PCR reactions using 1 ng (lanes 1,4), 2 ng (lanes 2,5), or 4 ng (lanes 3,6) of polyA⁺ RNA purified from v16;P[PSI] (lanes 1–3) or v16;P[PSIΔAB] males (lanes 4–6) and PCR primers specific for the indicated cDNA were resolved on 5% denaturing polyacrylamide gels and revealed by autoradiography. (D) Quantitative RT–PCR experiments were performed with (+) or without (−) reverse transcription step using 4 ng of polyA⁺ RNA and PCR primers specific for kettin (giant muscle protein, titin family), GABA-R (receptor for the inhibitory neurotransmitter GABA, Rdl subunit), or courtless cDNA (ubiquitin-conjugating enzyme, see Discussion). PCR products were analyzed as in C.