Fig. 2. Expression of
iff-1 and
iff-2. (A) The
iff-1 mRNA accumulation during development. RNA was prepared from wild-type hermaphrodite populations synchronized at each of the five different developmental stages (four larval and adult stages).
yk445a8 and
yk82f11 cDNA clones were used as probes to detect the
iff-1 and the
iff-2 transcripts, respectively. For each of
iff-1 and
iff-2 probes, we detected only a single band on the blot, which is shown here. A transcript of a ribosomal protein gene,
rpp-1, was used as a loading control. (B) Germline specific expression of the
iff-1 mRNA. Larval (top two panels) and adult (bottom two panels) hermaphrodites subjected to in situ hybridization using the
iff-1 cDNA clone,
yk445a8, as a probe. Top two panels are hermaphrodites at larval stages L1-L2 and L3-L4, respectively. Bottom two panels show the magnified images of the adult hermaphrodite gonad stained with the
iff-1 probe (upper) and DAPI (lower). The
iff-1 mRNA is abundant in the distal region of the gonad, with most intense staining roughly corresponding to the transition zone. Arrow, distal tip of the gonad; arrowhead, gonadal loop. Scale bars, 100 μm. (C) Western blot analysis using affinity-purified anti-IFF-1 antibodies and extracts from wild-type N2,
iff-1(
tm483),
glp-4(
bn2), and
gld-2(
q497)
gld-1(
q485) adult hermaphrodites. Protein extracts from 100 hermaphrodites of each genotype were loaded in each lane. The positions of IFF-1 and non-specific proteins are indicated by an arrowhead and asterisks, respectively. IFF-1 was not detected in the
iff-1 mutant, or in the
glp-4 mutant, which has very small gonads. Two bands were detected at the expected size of the IFF-1 protein (arrows) in the
gld-2 gld-1 double mutant, which has a number of proliferative germ nuclei in the gonad. The upper band may be the hypusinated IFF-1 protein. Size markers are shown on the right.