Extended Data Fig. 10: NFAT-RE-luciferase expression assay.
From: Bottom-up design of Ca2+ channels from defined selectivity filter geometry
a, Schematic diagram of the assay. Ca2+ influx through the designed channel in HEK293T cells would induce expression of the luciferase reporter through the Ca2+-dependent NFAT signaling pathway. b and c, Bar plots showing the average maximum luminescence intensity measured from cells (n = 3) expressing the wild-type CalC6_3 design (cyan), the E101L mutant (yellow), and the negative control (purple), respectively. The channel constructs contain a C-terminal mScarlet3 linked to the channel via the P2A self-cleaving sequence. The psPAX2 plasmid, which expresses proteins unrelated to Ca2+ flux or buffering, was used as the negative control. b, Raw luminescence intensity. c, Luminescence intensity normalized by mScarlet3 fluorescence. Error bars represent bootstrapped 95% confidence intervals around the mean.
