Fig. 4: Characterization of R-iLACCO variants in live mammalian cells.
From: Lactate biosensors for spectrally and spatially multiplexed fluorescence imaging
a Fluorescence images in HeLa cells. Scale bars, 10 μm. b ΔF/F in HeLa cells in (a). Parentheses represent number of cells investigated. c In situ l-lactate titration. n = 10, 18, and 18 cells for R-iLACCO1, R-iLACCO1.1, and R-iLACCO1.2, respectively (mean ± s.e.m.). d Time course of the fluorescence response in HeLa cells. n = 19, 21, and 21 cells for R-iLACCO1, R-iLACCO1.1, and R-iLACCO1.2, respectively (mean ± s.d.). One-way ANOVA followed by Tukey’s multiple comparison. *P < 0.0001. e Fluorescence traces (left) and ΔF/F (right) in response to blue-light illumination of HeLa cells. n = 17, 14, 11, and 14 cells for R-iLACCO1, R-iLACCO1.1, R-iLACCO1.2, and R-diLACCO1, respectively. Fluorescence traces represent mean ± s.e.m. f Photobleaching curves (left, mean ± s.e.m.) and integrated fluorescence (right) in HeLa cells. Inset shows the photobleaching curves in the first 10 s. n = 18, 20, 18, 19, and 17 cells for mApple, R-iLACCO1, R-iLACCO1.1, R-iLACCO1.2, and R-diLACCO1, respectively. One-way ANOVA followed by Tukey’s multiple comparison. *P < 0.0001. g Representative images (top) and fluorescence traces (middle) in HEK293T upon 5 mM d-glucose treatment after starvation. Box plots (bottom) show ΔF/F or ΔR/R for R-iLACCO variants and other biosensors. Parentheses represent number of cells investigated. Scale bars, 10 μm. h Representative images (top) and fluorescence traces (middle) in HeLa cells upon 1 μM AR-C155858 treatment. Box plots (bottom) show ΔF/F or ΔR/R for R-iLACCO variants and other biosensors. Parentheses represent number of cells investigated. Scale bars, 10 μm. i Representative images (top) and fluorescence traces (middle) in primary neurons upon 1 μM AR-C155858 treatment. Box plots (bottom) show ΔF/F or ΔR/R for R-iLACCO variants and other biosensors. Parentheses represent number of neurons investigated. Scale bars, 20 μm. j Representative images (top) and fluorescence traces (middle) in primary astrocytes upon 1 μM AR-C155858 treatment. Box plots (bottom) show ΔF/F or ΔR/R for R-iLACCO variants and other biosensors. Parentheses represent number of astrocytes investigated. Scale bars, 20 μm. Source data are provided as a Source Data file.
