Extended Data Fig. 1: RPA2 forms dynamic intracellular optoDroplets.
a, Comparison of blue light-inducible optoDroplet formation by a panel of DNA damage response proteins. FUSN fused to Cry2-mCherry and Cry2-mCherry E490A (Cry2olig) served as positive controls, GST fused to Cry2-mCherry and the empty Cry2-mCherry plasmid were included as negative controls. Representative stills from live-cell microscopy are shown. b, Accumulated optoDroplet intensity per nucleus of Cry2-mCherry constructs shown in (a) were analyzed and normalized to the average accumulated optoDroplet intensity of the corresponding dark condition. Two-way ANOVA with Šidák`s test, FUSN, Cry2oligo, RPA2, RNF8 **** p < 0.0001. c, Nuclear mean intensities of Cry2-mCherry in cells analyzed in (b). b-c, Averages and standard deviations are shown for n = 4 independent samples (cell number: emtpydark n1 = 604, n2 = 834, n3 = 785, n4 = 510; emptylight n1 = 587, n2 = 696, n3 = 781, n4 = 606; GSTdark n1 = 952, n2 = 966, n3 = 726, n4 = 727; GSTlight n1 = 871, n2 = 924, n3 = 1015, n4 = 863; FUSN dark n1 = 515, n2 = 652, n3 = 608, n4 = 571; FUSN light n1 = 750, n2 = 613, n3 = 787, n4 = 677; Cry2oligo dark n1 = 720, n2 = 941, n3 = 753, n4 = 828; Cry2oligo light n1 = 506, n2 = 889, n3 = 766, n4 = 952; RPA2dark n1 = 752, n2 = 864, n3 = 738, n4 = 647; RPA2light n1 = 811, n2 = 666, n3 = 965, n4 = 1028; RNF8dark n1 = 323, n2 = 437, n3 = 269, n4 = 493; RNF8light n1 = 235, n2 = 319, n3 = 356, n4 = 385; RNF168dark n1 = 459, n2 = 278, n3 = 532, n4 = 334; RNF168light n1 = 369, n2 = 506, n3 = 395, n4 = 424; CtIPdark n1 = 286, n2 = 217, n3 = 214, n4 = 246; CtIPlight n1 = 164, n2 = 213, n3 = 302, n4 = 262; RAD51dark n1 = 305, n2 = 245, n3 = 275, n4 = 124; RAD51light n1 = 256, n2 = 214, n3 = 243, n4 = 177). Scale bars 10 µm.
