DEV00916 Supplemental Figures

Files in this Data Supplement:

• Figure 1 -

  Fig. S1. Removal of GLP-1 activity increases GLD-1 accumulation in the distal end. This figure is similar to Fig. 3 in the text, however, wild-type and mutant hermaphrodite gonad arms are captured in the same field of view, and only the distal ends of the arms are shown. The top gonad arm is tumorous from a gld-2(q497) gld-1(q361); unc-32(e189) glp-1(q175) adult hermaphrodite with the distal end to the left (#). The bottom gonad arm is from a wild-type adult hermaphrodite and the distal end is to the right (*). Each panel is of the same two gonad arms, with the top panel stained with DAPI (blue), the middle panel showing proliferative germ cells (green) indicated by anti-REC-8 staining under our fixation conditions, and the bottom panel with GLD-1-specific antibodies (red). Wild-type animals have low GLD-1 levels in the most distal end while gld-2(q497) gld-1(q361); unc-32(e189) glp-1(q175) animals, which lack GLP-1/Notch signaling, have roughly equivalent GLD-1 accumulation levels in the very distal end and in regions immediately proximal where high levels are normally seen. Scale bar: 20 mm.

• Figure 2 -

  Fig. S2. GLP-1/Notch signaling inhibits GLD-1 accumulation. The distal portion of a tumorous gld-2(q497) gld-1(q361); unc-24(e138) lag-1(q426) hermaphrodite adult gonad arm (one day past L4 at 20�C) stained with DAPI (blue; top), REC-8 specific antibodies (green; middle) and GLD-1 specific antibodies (red; bottom). Distal is to the left. GLD-1 levels are higher in the most distal end than in wild-type animals, showing that a reduction in LAG-1 activity allows for increased accumulation of GLD-1 in the most distal end. Many of the germline nuclei are larger than in wild-type animals, the reason for this is currently unknown. Although the pattern of GLD-1 accumulation is similar to when GLP-1 activity is removed (Fig. 3; Supplemental Fig. S1), the overall levels of GLD-1 were somewhat lower. Scale bar: 20 mm.

• Figure 3 -

  Fig. S3. Males lacking GLD-1 and NOS-3 activity do not have tumorous germlines. gld-1(q485); nos-3(oz231) young adult males were dissected and stained with DAPI (blue) and REC-8-specific antibodies(green). Entry into meiotic prophase, meiotic progression and gametogenesis appears wild type. While 31% (n=36) of the males analyzed were indistinguishable from wild type (not shown), 69% of the males contained a small zone of proximal proliferation (shown in this figure). Scale bar: 20 mm.

• Figure 4 -

  Fig. S4.gld-2; nos-3 tumorous phenotype is epistatic to glp-1. Dissected hermaphrodite gonad arm from gld-2(q497); nos-3(oz231); unc-32(e189) glp-1(q175) young adult grown at 20�C stained with DAPI (blue) and phospho-histone (H3)-specific antibodies (red), which stain mitotic M-phase nuclei. Distal is to the left and marked with (*). These animals have tumorous, rather than Glp, germlines demonstrating that the gld-2; nos-3 tumorous phenotype is epistatic to glp-1. Scale bar: 20 mm.

• Figure 5 -

  Fig. S5.gld-2 and nos-3 promote GLD-1 accumulation. This figure is similar to Fig. 5B,E, except that the gonad arms are shown in the same field of view. Wild-type and tumorous animals were dissected, fixed and stained together. Wild-type and gld-2(q497); nos-3(oz231) arms were differentiated based on the tumorous phenotype. # marks the distal ends of the wild-type arms and *, the distal ends of the gld-1(q497); nos-3(oz231) arms. Animals were stained with DAPI (blue) and GLD-1-specific antibodies (red). GLD-1 accumulation can only be seen in the wild-type animals. Scale bar: 20 mm.