Germline P Granules Are Liquid Droplets That Localize by Controlled
Dissolution/Condensation
Clifford P. Brangwynne, Christian R. Eckmann, David S. Courson, Agata Rybarska,
Carsten Hoege, Jöbin Gharakhani, Frank Jülicher, Anthony A. Hyman
Supporting Online Material
This supplement contains:
Materials and Methods
SOM Text
Figs. S1 to S8
References
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Other Supporting Online Material for this manuscript includes the following: (available at www.sciencemag.org/cgi/content/full/1172046/DC1)
Movies S1 to S12
Movie s1
Movie shows P granule localization in GFP::PGL-1 embryo (same embryo as in Fig.
1A). DIC images have been colored red and overlaid over GFP::PGL-1 image
(green). As with all
Movie s2
Movie shows maximum intensity projection of a confocal z-stack of a GFP::PGL-1
embryo during symmetry breaking. Time shown is relative to pnm.
Movie s3
Movie shows an example of a GFP::PGL-1 embryo treated with spd-5(RNAi) for >24
hrs to delay symmetry breaking (same embryo as in Fig. 2D). Time = 0 defined as
first frame.
Movie s4
Movie shows mCherry::PAR-2/GFP::PGL-1 embryo treated with spd-5(RNAi) for
>24 hrs to delay symmetry breaking (same embryo as in Fig. S5). P granules appear
to completely dissolve prior to re-condensation upon the appearance of a cortical
PAR-2 domain. Time = 0 defined as first frame.
Movie s5
Movie shows GFP::MEX-5 embryo. At the beginning of the movie, the concentration
of MEX-5 is high throughout the embryo. However, the concentration in the posterior
soon decreases, and a significant gradient across the AP axis becomes apparent. Time
shown is relative to pnm.
Movie s6
Movie shows GFP::PGL-1 embryo treated with par-1(RNAi) for 24 hrs. P granules
dissolve throughout the embryo. Time shown is relative to pnm.
Movie s7
Movie shows syncytial germ cell nucleus covered in P-granules in the germ line of a
GFP::PGL-1 worm. The germ line has been dissected and squashed to induce
shearing forces (same nucleus as in Fig. 3B).
Movie s8
Movie shows syncytial germ cell nuclei covered in P granules in the germ line of a
GFP::PGL-1 worm. The germ line has been dissected and squashed. P granules
appear to drip off of the nuclei, fuse, and round up.
Movie s9
Movie shows syncytial germ cell nuclei covered in P granules in the germ line of a
GFP::PGL-1 worm. The germ line has been dissected and squashed. P granules
appear to de-wet from the nuclei, and can be seen to drip off of several nuclei.
Movie s10
Movie is a maximal intensity projection of a confocal stack of a GFP::PGL-1 embryo
treated with spd-5(RNAi) for >24 hrs to delay symmetry breaking, in which P
granules re-condense in the middle of the embryo (observed in <5% of embryos).
Note that at the beginning of this movie, P granules have already dissolved almost
completely, except toward the middle of the embryo. Time = 0 defined as first frame.
Movie s11
Movie shows a mCherry::PAR-2/GFP::PGL-1 embryo treated with spd-5(RNAi) for
>24 hrs to delay symmetry breaking (same as in Fig. S6). P granules can be seen in
the vicinity of each of the PAR-2 cortical domains, before one of the domains grows
larger and P-granules begin to condense there. Note also that P granule fusion events
can be seen towards the end of the movie. Time = 0 defined as first frame.
Movie s12
Movie shows nuclear associated P granules in an 8-cell GFP::GLH-1 embryo. Half of
one P granule is bleached. The fluorescence recovers within ~10 seconds.
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