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. 2005 Dec 1;25(1):88–96. doi: 10.1038/sj.emboj.7600901

Figure 3.

Figure 3

LIP-1 protein in the proximal mitotic region and FBF repression of lip-1 expression. (A–I, K) Adult germ lines dissected from animals of genotype, as labeled. Arrowhead, distal end; TZ, transition zone; A–F, staining with LIP-1-specific antibody; G–I, K, staining with antisense probe specific for lip-1 mRNA. Images in panels B–D were treated identically; same for panels E and F and for panels H and I. Tum(+FBF), gld-3 nos-3; Tum(−FBF), fbf-1 fbf-2 gld-3 nos-3. (A) LIP-1 protein in wild-type germ line. Upper image, DAPI staining used to assess nuclear morphology in the mitotic region, transition zone (TZ), and pachytene region; lower image, same germ line stained with LIP-1 antibody. LIP-1 is membrane associated and punctate, as shown previously (Hajnal and Berset, 2002). (B) LIP-1 protein in the mitotic region and transition zone of wild-type germ line. LIP-1 staining is membrane associated and punctate in proximal-most cells of the mitotic region and in transition zone (TZ). Blurry staining in cytoplasm is background (see C). (C) Background staining of LIP-1 antibody in germ line dissected from lip-1(zh15) deletion mutant. (D) Distal end of Tum(+FBF) germ line. Only background staining was detected (compare to C). (E) Complete Tum(+FBF) germ line. Staining with LIP-1 antibody is low relative to Tum(−FBF) and likely to be background. (F) Complete Tum(−FBF) germ line. LIP-1 staining is bright compared to Tum(+FBF). Also, LIP-1 is membrane associated and punctate throughout the germ line. (G) lip-1 mRNA in wild-type germ line. Distribution is similar to that of LIP-1 protein, but mRNA is not detected in the mitotic region, presumably owing to a lack of sensitivity. (H, I) In Tum(+FBF) germ lines, lip-1 mRNA level is lower than in Tum(−FBF). (J) Semiquantitative RT–PCR to compare abundance of lip-1 and unc-54 mRNAs in animals of genotype, as labeled. PCR cycles set within the linear range and amounts normalized to unc-54 product in wild type. (K) fbf-1 single mutant germ line. lip-1 mRNA is present in distal-most germ cells (bracket), which are being signaled by GLP-1/Notch signaling from the DTC.