Skip to main content
. Author manuscript; available in PMC: 2013 Aug 28.
Published in final edited form as: J Neurosci. 2013 Feb 27;33(9):4118–4127. doi: 10.1523/JNEUROSCI.4187-12.2013

Figure 6. Unbinding of naltrexamine alexa594 was not affected by previous agonist exposure.

Figure 6

A, Binding kinetics of the antagonist ntx A594 were measured by applying ntx A594 (100 nM, 5 min) and imaging binding and washout relative to M1–488 intensity (R/G). Binding and unbinding was plotted and fit with a single exponential decay formula (tau association= 65.8±5.5 sec, tau off =184±6.1 sec, n=4 each, average±sem). B, Preincubation of cells with the irreversible antagonist β-CNA (1 µM, 5 min) prevented all binding when measured by the fluorescence intensity immediately after washout of ntx A594 (300 nM, 90 sec) verifying the specificity of binding (n=2 ctrl, n=4 β-CNA mean±sd). C, Ntx A594 (300 nM, 90 sec) was applied and unbinding of ntx A594 was imaged from cells either untreated (black) or pre-treated with ME (30 µM, 2 hr) (red) n=5 each, average±sem.