Figure 5.

Masked mRNAs and speckle markers enter the cytoplasm during the first division and Mago is required for their asymmetrically redistributed. a-c, 25mer biotinylated probes directed against SPDS transcripts were hybridized and detected using avidin conjugated Fluorescein (green) and counter stained with DAPI (blue). Bar = 10 μm. a, Microspore prior to the prothallial division. b, Microspore during the prothallial division. c, Microspore just after completion of the prothallial division. The prothallial nucleus is denoted by "p" in b and c. d-u, "sp" denotes spermatogenous cells, "jk" denotes jacket cells, arrowheads mark foci of labeling within spermatogenous cells and arrows mark foci of labeling within jacket cells. d-f, Microspore fixed at 4 hours of development and probed with short FISH probes for masked centrin mRNA (red). d, DAPI (blue), e phase contrast, and f centrin FISH probes detected with TexasRed conjugated antibody (red). g-i, microspore fixed at 4 hours of development and probed with 25mer FISH probes for masked SPDS mRNA (green). g, DAPI (blue), h phase contrast, and i SPDS FISH probes detected with Fluorescein conjugated antibody (green). j-k, 4G3 labeling of U2B" protein (red) at (j) 2 hours, (k) 4 hours, and (l) 5 hours of development. Arrowheads denote cytoplasmic masked mRNA (f and i) and U2B" (j-l). m-o, Microspore subjected to Mv-Mago RNAi and fixed after 4 hours of development. (m) DAPI (blue), (n) phase contrast, (o) masked centrin transcripts (red) detected with 25mer biotinylated FISH probes. Arrows denote masked transcripts within jacket cells. Boxed regions enlarged in insets. Spermatogenous (sp) and jacket cells (jk) labeled in n, q and t. p-r, Microspore subjected to Mv-Mago RNAi and fixed after 4 hours of development. (p) DAPI (blue), (q) phase contrast, (r) masked SPDS transcripts (green) detected with 25mer biotinylated FISH probes. Arrows denote masked transcripts within jacket cells. Boxed regions enlarged in insets. s-u, Microspores subjected to Mv-Mago RNAi and fixed after 5 hours of development. (s) DAPI (blue), (t) phase contrast, (u) 4G3 labeling of U2B" (red). Arrow denotes U2B" within jacket cell. Bar = 25 μm.

Boothby and Wolniak BMC Cell Biology 2011 12:45   doi:10.1186/1471-2121-12-45
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