Figure S4.
mRNA promotes LLPS and modulates the material properties of PGL condensates, related toFig. 6. (A–C) Purified PGL-1/-3/SEPA-1 proteins at 0.2 μM for each fail to undergo LLPS, while a few small condensates are formed in the presence of 10 ng/μl mRNA. C shows the average number of condensates in A and B. Data are shown as mean ± SEM (n = 3; n refers to the number of randomly selected fields (220 × 166 µm) for each reaction). (D–F) Adding mRNA enlarges the size of condensates formed by purified PGL-1/-3/SEPA-1 proteins (1 µM for each protein). (F) shows column scatter charts of the diameters of condensates in D and E. Data are shown as mean ± SEM of condensates combined from three fields (220 × 166 µm) for each reaction (n = 106 for no mRNA and n = 154 for 10 ng/μl mRNA; n refers to the total number of condensates analyzed for each reaction). Two-tailed, unpaired t test result: ***P < 0.001. (G–I) Adding mRNA slightly increases the partitioning of TagRFP::IFE-1 into PGL-1/-3/SEPA-1 condensates (3 µM for each protein). (I) shows the mean TagRFP::IFE-1 fluorescence intensity per AU in condensates formed by 3 µM PGL-1/-3/SEPA-1/TagRFP::IFE-1 with or without the addition of 30 ng/μl mRNA. Data are shown as mean ± SEM (n = 27; n refers to the number of condensates analyzed for each reaction). Two-tailed, unpaired t test result: ***P < 0.001. (J–L) DIC images showing that addition of mRNA enlarges the size of condensates formed by PGL-1/-3/SEPA-1/GFP::EPG-2 (3 µM for each). (L) shows column scatter charts of the diameters of condensates in J and K. Data are shown as mean ± SEM of condensates combined from three fields (128 × 90 µm) for each reaction (n = 160 for no mRNA and n = 139 for 30 ng/μl mRNA; n refers to the total number of condensates analyzed for each reaction). Two-tailed, unpaired t test result: ***P < 0.001. (M) Protein gels for the inputs used in the in vitro LLPS assay (asterisks indicate the corresponding bands). Scale bars: 20 µm for A, B, D, E, G, H, J, and K; 5 µm for enlarged images in A, B, D, E, J, and K. Source data are available for this figure: SourceData FS4.

mRNA promotes LLPS and modulates the material properties of PGL condensates, related to Fig. 6 . (A–C) Purified PGL-1/-3/SEPA-1 proteins at 0.2 μM for each fail to undergo LLPS, while a few small condensates are formed in the presence of 10 ng/μl mRNA. C shows the average number of condensates in A and B. Data are shown as mean ± SEM (n = 3; n refers to the number of randomly selected fields (220 × 166 µm) for each reaction). (D–F) Adding mRNA enlarges the size of condensates formed by purified PGL-1/-3/SEPA-1 proteins (1 µM for each protein). (F) shows column scatter charts of the diameters of condensates in D and E. Data are shown as mean ± SEM of condensates combined from three fields (220 × 166 µm) for each reaction (n = 106 for no mRNA and n = 154 for 10 ng/μl mRNA; n refers to the total number of condensates analyzed for each reaction). Two-tailed, unpaired t test result: ***P < 0.001. (G–I) Adding mRNA slightly increases the partitioning of TagRFP::IFE-1 into PGL-1/-3/SEPA-1 condensates (3 µM for each protein). (I) shows the mean TagRFP::IFE-1 fluorescence intensity per AU in condensates formed by 3 µM PGL-1/-3/SEPA-1/TagRFP::IFE-1 with or without the addition of 30 ng/μl mRNA. Data are shown as mean ± SEM (n = 27; n refers to the number of condensates analyzed for each reaction). Two-tailed, unpaired t test result: ***P < 0.001. (J–L) DIC images showing that addition of mRNA enlarges the size of condensates formed by PGL-1/-3/SEPA-1/GFP::EPG-2 (3 µM for each). (L) shows column scatter charts of the diameters of condensates in J and K. Data are shown as mean ± SEM of condensates combined from three fields (128 × 90 µm) for each reaction (n = 160 for no mRNA and n = 139 for 30 ng/μl mRNA; n refers to the total number of condensates analyzed for each reaction). Two-tailed, unpaired t test result: ***P < 0.001. (M) Protein gels for the inputs used in the in vitro LLPS assay (asterisks indicate the corresponding bands). Scale bars: 20 µm for A, B, D, E, G, H, J, and K; 5 µm for enlarged images in A, B, D, E, J, and K. Source data are available for this figure: SourceData FS4.

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