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[Preprint]. 2024 Oct 2:2024.09.30.615856. [Version 1] doi: 10.1101/2024.09.30.615856

Figure 2. Tuning the NLS of ERK-KTR.

Figure 2.

(A) Amino acid sequence alignments of ERK-KTR sensor domains, showing the kinase docking site, bNLS, (pink, asterisk) PKA phosphorylation sites, and NES, with predicted NLS strength scores listed to the right. Bar, 50 μm. (B) Representative live-cell fluorescence images of HEK293 cells expressing H2B-mTagBFP2, mNeonGreen, and variants of ERK-KTR/tdCherry proteins. -EGF refers to fluorescence images taken prior to the addition of EGF, while +EGF refers to fluorescence micrographs taken 15 minutes after the addition of 10 nM EGF. (C) Bar graph showing C/N ratios of eERK-KTR1.1, 1.2, and 1.3 /tdCherry1, following 15 minutes incubation in 0, 0.3, 3, or 10 nM EGF. (D) C/N ratios of eERK-KTR1.2/mScarlet (gold) or eERK-KTR1.2/mScarlet/BCR (brown), after 15 minutes incubation in 0, 0.1, 0.3, 1, or 10 uM Fsk. Data are from a minimum of three independent biological replicates. p values were calculated by two-way ANOVA.