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. Author manuscript; available in PMC: 2021 Jul 1.
Published in final edited form as: Nat Chem Biol. 2020 Sep 28;17(1):39–46. doi: 10.1038/s41589-020-00660-y

Extended Data Fig. 4 |. ExRai-AKAR2 is a more sensitive FACS probe than ExRai-AKAR1.

Extended Data Fig. 4 |

HEK293T cells transfected with ExRai-AKAR1 were analyzed via flow cytometry before and after stimulation with 50 μM Fsk and 100 μM IBMX as described in the Methods. a, Contour plot showing the 488 nmand 405 nm-excited fluorescence intensities of transfected cells without (teal) and with (green) Fsk/IBMX stimulation. b, Frequency distribution of 488 nm/405 nm excitation ratio illustrating the population shift caused by Fsk/IBMX stimulation (****P < 0.0001; Kolmogorov-Smirnov test). Data are representative of 3 independent experiments. Overlaid gray and black dashed lines depict the frequency distributions for ExRai-AKAR2 transfected cell populations before and after Fsk/IBMX treatment, respectively (redrawn from Fig. 3a). c, Table summarizing the input values and results of the sensitivity index (SI) calculation (see Methods). ExRai-AKAR2 shows 2-fold higher sensitivity compared with ExRai-AKAR1.