Figure 3. The ectopic somatic cells in the ER-stressed gonad of gld-1-deficient animals are germ cell-derived differentiated somatic cells.

(A) Representative fluorescence micrographs (x400) of somatic differentiation markers expressed in the gonads of gld-1; ced-3-deficient animals on day-4 of adulthood. gld-1 RNAi was used to sensitize the germline for transdifferentiation. ced-3 RNAi was used to prevent the clearance of the ectopic somatic cells from the gonad. Punc-119::gfp is a neuronal marker, Pmyo-2::gfp is a pharyngeal muscle marker. Pelt-2::NLS::gfp is an intestinal marker. ER stress was induced by tfg-1 RNAi or by an xbp-1 mutation. Asterisks mark Student's T-test values of p < 0.001 compared to non-stressed conditions. At least 40 animals were analyzed per genotype. (B) Representative fluorescence micrographs (x400) of Punc-119::gfp in the gonads of ced-3(n1286) day-4 animals treated with gld-1 RNAi. ER stress was induced chemically with tunicamycin and compared to DMSO treatment. (C) Representative fluorescence micrographs of DAPI-stained nuclei of ER-stressed day 4 adults treated with a mixture of gld-1, ced-3 and tfg-1 RNAi. Accumulation of abnormal somatic-like nuclei was detected in germ cell(+) animals and not in germ cell(−) glp-1(−) mutants. At least 50 gonads were analyzed per genotype. Asterisk marks Student's T-test values of p < 0.001. See Figure 3—figure supplement 1 for co-localization of the somatic marker expressing cells and the cells with the large nuclei and/or the cell under engulfment.

DOI: http://dx.doi.org/10.7554/eLife.08005.008

Figure 3—figure supplement 1. The ectopic somatic cells in the tumorous gonad have large nuclei and are engulfed by the surrounding gonad.

(A) Day 4 Punc-119::gfp transgenic animals treated with a mixture of gld-1, ced-3 and tfg-1 RNAi were stained with the nuclear dye Hoechst. Hoecst staining and GFP expression were individually captured and used to assess co-localization between the pattern of the GFP-expressing cells in the gonad and the cells harboring ectopically large nuclei. (B) Day 3 transgenic animals co-expressing the somatic marker Pelt-2::NLS::GFP and the engulfment marker Plim-7::ced-1::gfp were treated with a mixture of gld-1 and tfg-1 RNAi. Solid arrow indicates an engulfed cell expressing the somatic marker, demonstrating that the ectopic somatic cells in the tumorous gonad are engulfed and removed by the surrounding cells. Dashed arrow indicates an engulfed cell which does not express the somatic marker.

DOI: http://dx.doi.org/10.7554/eLife.08005.009