Fig. 5
Loss of SUP-46 disrupts P granules, causes ectopic spermatogenesis, and increases spermatogenesis transcripts. a 67% (580/872) of spermatogenesis transcripts were increased (P = 2.4 × 10–482) and < 1% (2/872) decreased (P = 4.0 × 10–14) in the sup-46 mutant. b In contrast, 51% (180/353) of oogenesis transcripts were decreased (P = 3.3 × 10–156) and none were increased (P = 2.5 × 10–12) in the sup-46 mutant. c 3% (14/460) of hermaphrodite soma transcripts were increased (P = 7.9 × 10–5) and 6% (26/460) decreased (P = 0.06) in the sup-46 mutant. d 4% (16/433) of male soma transcripts were increased (P = 0.001) and 4% (19/433) decreased (P = 0.42) in the sup-46 mutant. e 71% (414/580) of the sperm transcripts that were increased in the sup-46 mutant were also increased in P granule(RNAi) (P = 1.1 × 10–353). f In contrast, none of the oocyte transcripts that were down in the sup-46 mutant were also down in P granule(RNAi) (P = 5.0 × 10–4). g PGL-1,3 were clustered in perinuclear granules in the germline of wildtype males, but diffusely distributed in glh-1 and sup-46 mutants. h glh-1 and sup-46 mutants had ectopic sperm (arrow) interspersed with pachytene nuclei in the middle region of the germline. The proximal germline, to which sperm are normally confined, is indicated. i Immunolocalization with the sperm-specific antibody, anti-SP56, confirms identification of sperm in the middle region of a sup-46 mutant male. Regions containing sperm are outlined with a hatched line. The inset (enlarged) shows sperm marked by SP56, and is a different focal plane of the area marked by the asterisk. In wildtype, sperm were restricted to the proximal region, while the sup-46 mutant had sperm extending as far back as the loop region. (a–f) Venn diagram circle sizes and overlaps are proportional to the number of genes in each class. Sperm and oocyte transcripts are the overlapping sets of [75] and [74], soma transcripts are from [75], and P granule(RNAi) are from [76]. P values were calculated using the online hypergeometric distribution calculator, (http://systems.crump.ucla.edu/hypergeometric/index.php). (g–i) Germlines from male worms were fixed and stained with antibodies and PicoGreen (DNA). Scale bars represent 15 μm. Images are of a single confocal optical plane