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. 2012 Apr 11;32(15):5200–5208. doi: 10.1523/JNEUROSCI.6347-11.2012

Figure 4.

Figure 4.

Comparison between sustained and simultaneous application of AEA-induced potentiation of GlyRs. A, Potentiation of IGly induced by 5 min sustained incubation of THC (1 μm)/AEA (10 μm) in HEK-293 cells expressing α1, α2 and α3 GlyRs. B, Potentiation of IGly induced by simultaneous application of THC (1 μm)/AEA (10 μm) in HEK-293 cells expressing α1, α2 and α3 GlyRs. C, Current trace showing potentiation of IGly induced by simultaneous application of AEA or 5 min sustained incubation of 10 μm AEA in HEK-293 cells expressing the S296A mutant and wild-type α1 GlyRs. The solid bars on the top of traces indicate the time of drug application. D, Quantification of data in experiments shown in C. ***p < 0.001, unpaired t test. Note that the S296A mutation completely abolished the additional potentiation of IGly induced by sustained AEA incubation but not by simultaneously coapplied AEA. E, Amino acid alignment of the TM2 region flanking G254 (α1) or equivalent residues in the α2 and α3 subunits. F, The average percentage potentiation induced by sustained AEA application in HEK-293 cells expressing the G254A mutant and wild-type α1 GlyRs.