Supplementary Figure 2
From: A gel phase promotes condensation of liquid P granules in Caenorhabditis elegans embryos
Effect of temperature and dilution of MEG and PGL phases. a Time-lapse photomicrographs of a 2-cell embryo expressing MEG-3::GFP and PGL-3::mCherry fusions and cultured at 20 °C, up-shifted to 34 °C for 15 minutes (same experiment as in Fig. 2, but longer period at 34 °C). MEG-3 remains in granules throughout. PGL-3 disperses at 34 °C. Scale bar is 10 μm. b Time-lapse photomicrographs of a 2-cell embryos expressing MEG-3::mCherry and PGL-1::GFP fusion cultured at 20 °C and then up-shifted to 34 °C for 1 minute. MEG-3 remains in granules throughout. PGL-1 disperses at 34 °C. Scale bar is 10 μm. c Extruded MEG-3::GFP granules over time. Time-lapse photomicrographs of MEG-3::GFP P granules before and after extrusion into egg buffer (top panels). Time after extrusion is indicated in each image. Scale bar is 10 μm. Graph (bottom panel) showing the fraction of MEG-3::GFP retained in the granular phase over time after extrusion. Total GFP fluorescence from granules was measured 1 min after extrusion (IB) and at times indicated after extrusion (IA) and used to calculate a fluorescence ratio (IA/IB) (n = 10). d Time-lapse photomicrographs of a 4-cell embryo expressing MEG-3::mCherry or 2-cell embryos expressing PGL-1::GFP before and 10 s after laser puncture of the eggshell (top panels). Laser puncture causes the contents of the posterior blastomere to spill onto the slide mixing with the aqueous buffer. MEG-3::mCherry persists in the granule phase, whereas PGL-1 becomes dispersed. Scale bar is 10 μm. Graph (bottom panel) showing the fraction of MEG-3::mCherry (n = 5) or PGL-1::GFP (n = 3) retained in the granular phase after extrusion. Total GFP fluorescence from granules was measured before laser puncture (IB) and after laser puncture (IA) and used to calculate a fluorescence ratio (IA/IB). Means are indicated along with error bars representing ± SD.
