Expression of DXZ4 in male and female macaques. (a) Schematic representation of a single DXZ4 monomer with positions in kilobases indicated. Below the monomer the blue bars indicate the relative positions of regions of macaque DXZ4 assessed by RT-PCR with the coordinates of the fragment indicated. Each blue bar points to the corresponding RT-PCR data obtained from male and female rhesus macaque cDNA. Samples indicate a no template control (Water) and cDNA prepared form total RNA with (+RT) and without (-RT) including reverse transcriptase enzyme. (b) Quantitative RT-PCR (qRT-PCR) analysis of DXZ4 expression in male and female rhesus macaque cDNA. Data represent the mean of four independent triplicate qRT-PCR analyses and the black bar indicates the standard error. Expression levels are represented as a percentage of GAPDH levels. (c) Strand-specific RT-PCR of DXZ4. The locations of primers used to prime anti-sense or sense cDNA synthesis are indicated by the right and left-facing arrow-heads and the precise location in DXZ4 is given by the coordinates. The red box represents the region assessed by PCR for anti-sense transcripts and the blue box for sense transcripts. Below the schematic map is the RT-PCR data as an ethidium bromide stained agarose gel. Samples include a no template control (Water), strand-specific primer with reverse transcriptase (+RT), no strand-specific primer with reverse transcriptase (No oligo +RT) and no primer with no reverse transcriptase (-RT). (d) RNA FISH analysis of DXZ4 expression in male and female rhesus macaque nuclei. Nuclei are counterstained with DAPI (blue). DXZ4 expression (red) is indicated by the white arrowheads. XIST RNA expression (green) is indicated by the white arrows. (e) Quantitative analysis of DXZ4 expression from the Xa and Xi in male and female rhesus macaque (R. Macaque) and pig-tailed macaque (P-T. Macaque) (n = 50). (f) RNA FISH analysis of MIC2 expression (red) relative to XIST (green) in male and female rhesus macaque nuclei.