Figure 1.
PGL granules formed under heat-stress conditions contain proteins involved in translation. (A and B) IFE-1::GFP forms a few granules in the germ precursor cells (Z2 and Z3, highlighted in dashed circles), and is diffusely localized in the cytoplasm of somatic cells in wild-type (WT) embryos at 20°C. Embryos at the ∼100-cell stage and the comma stage are shown in A and B, respectively. (C and D) Temporal accumulation of IFE-1::GFP granules in somatic cells in WT embryos laid at 26°C. IFE-1::GFP forms a few tiny granules at the ∼100-cell stage (C), and forms a large number of granules at the comma stage (D). (E and F) IFE-1::GFP granules are absent from somatic cells in sepa-1(bp1726) (E) and pgl-1(RNAi) (F) embryos at the comma stage at 26°C. In pgl-1(RNAi) embryos, the number of IFE-1::GFP granules is also reduced in two-germ precursor cells (highlighted in dashed circles). Maximum-intensity projections of Z-stack confocal images are shown in A–F. (G) Quantification of the number of IFE-1::GFP granules in somatic cells per embryo at the ∼100-cell and comma stage at 20 and 26°C. Data are shown as mean ± SEM (n = 3; n refers to the number of embryos analyzed for each genotype). Two-tailed, unpaired t-test results: ***P < 0.001. (H) IFE-1::GFP granules are colocalized with PGL granules, labeled by TagRFP::SEPA-1, in WT embryos laid at 26°C. PGL granules contain oocyte-derived PGL-1/-3 and the zygotically synthesized SEPA-1. PGL-1/-3 are present in both P granules and PGL granules, while SEPA-1 is present only in PGL granules. (I) Quantification of the percentage (%) of PGL granules positive for IFE-1::GFP in WT and atg-3 mutant embryos. Data are shown as mean ± SEM (n = 3; n refers to the number of embryos analyzed for each genotype or condition). Two-tailed, unpaired t-test results: **P < 0.01, ***P < 0.001. (J and K) Purified TagRFP::IFE-1 protein is partitioned into PGL-1/-3/SEPA-1 condensates (3 µM for each protein) in in vitro LLPS assays. J is the DIC image of K. His-tagged PGL-1 and PGL-3, His- and TagRFP-tagged IFE-1 isoform b, and His- and MBP-tagged SEPA-1 were used for LLPS in this study unless otherwise noted. Scale bars: 5 µm for A–F and H; 2 µm for enlarged images in H; 20 µm for J and K.

PGL granules formed under heat-stress conditions contain proteins involved in translation. (A and B) IFE-1::GFP forms a few granules in the germ precursor cells (Z2 and Z3, highlighted in dashed circles), and is diffusely localized in the cytoplasm of somatic cells in wild-type (WT) embryos at 20°C. Embryos at the ∼100-cell stage and the comma stage are shown in A and B, respectively. (C and D) Temporal accumulation of IFE-1::GFP granules in somatic cells in WT embryos laid at 26°C. IFE-1::GFP forms a few tiny granules at the ∼100-cell stage (C), and forms a large number of granules at the comma stage (D). (E and F) IFE-1::GFP granules are absent from somatic cells in sepa-1(bp1726) (E) and pgl-1(RNAi) (F) embryos at the comma stage at 26°C. In pgl-1(RNAi) embryos, the number of IFE-1::GFP granules is also reduced in two-germ precursor cells (highlighted in dashed circles). Maximum-intensity projections of Z-stack confocal images are shown in A–F. (G) Quantification of the number of IFE-1::GFP granules in somatic cells per embryo at the ∼100-cell and comma stage at 20 and 26°C. Data are shown as mean ± SEM (n = 3; n refers to the number of embryos analyzed for each genotype). Two-tailed, unpaired t-test results: ***P < 0.001. (H) IFE-1::GFP granules are colocalized with PGL granules, labeled by TagRFP::SEPA-1, in WT embryos laid at 26°C. PGL granules contain oocyte-derived PGL-1/-3 and the zygotically synthesized SEPA-1. PGL-1/-3 are present in both P granules and PGL granules, while SEPA-1 is present only in PGL granules. (I) Quantification of the percentage (%) of PGL granules positive for IFE-1::GFP in WT and atg-3 mutant embryos. Data are shown as mean ± SEM (n = 3; n refers to the number of embryos analyzed for each genotype or condition). Two-tailed, unpaired t-test results: **P < 0.01, ***P < 0.001. (J and K) Purified TagRFP::IFE-1 protein is partitioned into PGL-1/-3/SEPA-1 condensates (3 µM for each protein) in in vitro LLPS assays. J is the DIC image of K. His-tagged PGL-1 and PGL-3, His- and TagRFP-tagged IFE-1 isoform b, and His- and MBP-tagged SEPA-1 were used for LLPS in this study unless otherwise noted. Scale bars: 5 µm for A–F and H; 2 µm for enlarged images in H; 20 µm for J and K.

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