FISH analysis of satellite VI in the metaphase spreads of a male Indian muntjac. (a) A strong satellite VI DNA signal (red) was detected at the pericentric X region of chromosome X + 3 and 14 interstitial hybridization signals were observed after longer exposure. Inset: a clear satellite VI signal was observed at the Xp11.1 side of the X + 3 pericentromere after short exposure. (b) The G-banded like chromosomes are presented from the inverted DAPI image of panel (a). Red interstitial satellite VI signals were indicated by arrows. (c) All interstitial satellite VI fluorescence signals (red; indicated by arrows) are co-localized with satellite I (C5 as a probe; green). (d) Triple-color FISH study with biotinylated satellite I (presented as a pseudo-green color), Cy3 direct-labeled satellite II (Mmv-0.7 as a probe; presented as a pseudo-orange color) and digoxigenin-labeled satellite VI (presented as a pseudo-red color) cohybridized to the metaphase chromosomes. The satellite II signal is located in the centromeric region of the chromosome X + 3. The satellite I signal is located in both pericentromere regions of the X + 3. The satellite VI signal is located in the Xp11.1 of the X + 3 pericentromere region and is more distal to the centromere than the satellite I signal. The FISH image was captured under low exposure time (scale bar = 10 μm). (e) Schematic illustration of the chromosome X + 3 in the panel (d) which shows the organization of cervid satellite I, II, and VI in the centromeric and pericentromeric regions of chromosome X + 3. The exact boundary of each satellite DNA could not be exactly defined and was indicated by dotted lines.