Plan apo vc 100x 1.4 oil

For experiments examining Dcp1/Dcp2_ext recovery, condensates were formed with 40 μM Dcp1/Dcp2_ext. Experiments monitoring the effects of Edc3 were performed with 5 μM Dcp1/Dcp2 and 5 μM or 80 μM Edc3. For analysis of RNA recovery, condensates were formed with 40uM Dcp1/Dcp2_ext with or without Edc3 and FITC-29mer RNA was added. The concentration of labeled protein or RNA was 250 nM across all samples. Samples were imaged in a passivated glass bottom 384-well plate (Greiner Bio-One). Imaging was performed at room temperature using an inverted Nikon Ti microscope equipped with an Andor Borealis CSU-W1 spinning disk confocal, Plan Apo VC 100x/1.4 oil objective and Andor iXon Ultra DU888 EMCCD camera. For each photobleaching experiment, a rectangular region of interest (ROI) was drawn around single condensates and irradiated for 1.5s with 7mW power at 473 nm with a Vortran laser between the fifth and sixth acquired frame. For analysis, three ROIs were used corresponding to the bleached droplet, an unbleached droplet, and background. Analysis was performed using ImageJ. Recovery traces were obtained by performing a double normalization to account for photobleaching during image acquisition^{53 (link)}. Recovery t_1/2 and immobile fractions were determined from fits of single exponentials using Prism 8/9 (GraphPad) and are reported in Supplementary Table 2.