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Fig. 2. Localization patterns of xARH isoforms during oogenesis. Oocytes are oriented with animal pole on top. Sense probe served as the negative control. (A)
xARH localizes to the vegetal cortex (arrow). DIG labeled xARH probe covering the coding region was hybridized to bisected stage V oocytes. (B) xARH
localizes to the vegetal cortex of oocytes through the late pathway. Whole mount in situ analysis of stage II albino oocytes. In early (small) stage oocytes,
xARH distributes homogeneously while in late (large) stage oocytes, xARH localizes to the vegetal pole. Staining was also found within the animal hemisphere.
(C) Stage VI oocytes and 16-cell embryos were bisected into animal and vegetal halves and total RNA isolated. Isoforms of xARH localize to different domains
in oocytes and embryos. Northern blot analysis of RNA from total oocytes or embryos (T), animal (A) or vegetal (V) halves using the full-length xARH probe.
ODC served as a loading control. |
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Fig. 4. Expression of xARHa and b RNAs during embryogenesis. (A) Total RNA was isolated from different embryonic stages. 28s rRNA served as the loading
control. (B) Blastula stage embryos were hybridized with DIG-labeled xARH. Only the vegetal hemisphere is shown. The inset shows a section of the embryo
magnified to reveal the perinuclear localization pattern (arrow). Hybridization in each case was with probe covering the coding region of xARH. (C) Northern
blot showing expression of xARHa and b RNAs in adult tissues. Approximately 10 mg total RNA was loaded onto each lane. Full length xARHa was used as
the probe. xARH is expressed at high levels in ovary and at relatively low levels in liver and spleen. Some signal can also be detected in heart and testis. 18 s
rRNA served as the loading control. |
