Fig. 2 | Nature Communications

Fig. 2

From: Ratiometric Matryoshka biosensors from a nested cassette of green- and orange-emitting fluorescent proteins

Fig. 2

In vitro characterization of MatryoshCaMP6s and sfMatryoshCaMP6s and sfMatryoshCaMP6s-T78H. a Schematic representation of MatryoshCaMP6s sensors, composed of GO-Matryoshka (LSSmOrange sandwiched between the C and N termini of either EGFP, sfGFP or sfGFP-T78H) inserted between the M13 peptide and calmodulin domain. LE and LP represent flanking residues that connect the cpFP domains with the recognition domains. b Steady-state fluorescence spectra (λ exc 440 nm) of sensors titrated with free calcium (0, 0.02, 0.03, 0.06, 0.09, 0.13, 0.20, 0.31, 0.52, 1.2 and 37 µM). The FI change in the orange channel mainly derives from fluorescence bleed through. c Calcium-affinity titrations (R = FI510nm/FI570nm) corresponding to the spectra in b. Data were corrected for fluorescence bleed-through (factor 0.1; mean ± s.e.m. from biological replicates with n = 6 for MatryoshCaMP6s and sfMatryoshCaMP6s, n = 4 for sfMatryoshCaMP6s-T78H)

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