Fig. 5.

Characterization of cell surface and cytosolic iATPSnFR^1.0 in astrocytes and neurons. a A single plane from a confocal z-stack of a U373MG cell expressing cytoplasmic mCherry and membrane-displayed iATPSnFR^1.0. b Confocal images of U373MG cells expressing iATPSnFR^1.0 before ATP application (control) and during a 300 µM ATP application (+ATP), as well as the change in fluorescence (dF). c Confocal images of hippocampal neurons before ATP application (control) and during 1 mM ATP application, as well as the change in fluorescence (dF). d, e Dose–response curves for iATPSnFR^1.0 when displayed on the surface of U373MG astroglia and hippocampal neurons. f A confocal image of a U373MG astroglia cell transfected with iATPSnFR^1.0 before a 3 mM puff of ATP (control), during the puff (ATP), as well as the change in fluorescence (dF). An average trace from all cells with the s.e.m. is also shown. g Representative hippocampal astrocyte in acute brain slice expressing iATPSnFR^1.0 before the application of ATP (control), during the application of 3 mM ATP (ATP), and the dF. An average trace from all cells with the s.e.m. is also shown. h Scatter graph summary of the effect of 3 mM ATP on iATPSnFR^1.0 expressed in the cell types indicated. In panel (h), the circle represents the mean, the box the s.e.m., the whiskers the s.d., and the horizontal line the median. p Values are shown for statistical comparisons to the buffer puff control using un-paired Student’s t tests. Abbreviations: hip hippocampus, SC spinal cord, MSNs medium spiny neurons. Note broken y-axis. In other panels, the error bars represent the s.e.m. and in some cases are smaller than the symbols used for the mean. n numbers are provided in the figure panels and refer to the number of cells