Fig. 3

Characterization of Electra1, Electra2 in mammalian cell culture. (a) Blue-to-green fluorescence ratio in live HEK cells co-expressing mBlueberry2, EBFP2, mTagBFP2, Electra1, and Electra2 with EGFP via the P2A self-cleaving peptide (n = 835, 573, 1698, 1229, and 664 cells respectively, 4 independent transfections each; Kruskal–Wallis ANOVA, p-value = 0; post-hoc Kolmogorov-Smirnoff two sample p-values shown in graph; Supplementary Table S4). Imaging conditions for BFPs: 403 nm LED excitation, emission 456 nm, 0.91 mW/mm2 power. Box plots same as in Fig. 2b. (b) Normalized photobleaching curves for mBlueberry2, EBFP2, mTagBFP2, Electra1 and Electra2 expressed in HEK cells (n = 97, 81, 122, 105, 70 cells respectively, 4 independent transfections each; imaging conditions for BFPs: excitation 403 nm LED excitation, emission 456 nm, 0.91 mW/mm2 power). Dotted lines represent linear fit (decrease rate for EBFP2, mTagBFP2, Electra1, Electra2 in fluorescence %/sec: 8.33195e-4, 487.4e-4, 282e-4, 341.3e-4, respectively, fitting details in Supplementary Table S4). (c) Normalized photobleaching curves for mBlueberry2, EBFP2, mTagBFP2, Electra1 and Electra2 expressed in HEK cells (n = 63, 79, 137, 144, 90 respectively; 4 independent transfections each; imaging conditions for HEK cells: 403 nm LED excitation, 456 nm emission, 4.25 mW/mm2 power).