Table 4.

Diffusion analysis of synaptic and extrasynaptic TMD-βL variants and full-length GlyRβ subunits in rat spinal cord neurons

Parameter	β-Loop variant	Synaptic^⁸ Q1/Q2/Q3	Extrasynaptic^⁹ Q1/Q2/Q3
Explored area [10⁻³ µm²]^¹	TMD-βL-wt^³	0.67/1.23/3.35	0.82/1.63/4.96
	TMD-βL-HI^⁴	0.64/1.16/2.21	0.75/1.39/3.53
	TMD-βL-βL-ΔCore^⁵	1.86/4.60/11.42	1.51/4.63/12.77
	TMD-βL-D407P/F408G^⁶	1.97/6.58/16.56	2.33/7.13/17.92
	TMD-βL-geph⁻^⁷	4.27/11.50/23.03	3.86/11.49/24.85
D_eff [µm² * s⁻¹]^²	TMD-βL-wt^³	0.02/0.05/0.11	0.04/0.07/0.26
	TMD-βL-HI^⁴	0.03/0.06/0.12	0.04/0.08/0.30
	TMD-βL-ΔCore^⁵	0.05/0.12/0.30	0.06/0.21/0.57
	TMD-βL-D407P/F408G^⁶	0.05/0.15/0.40	0.10/0.32/0.73
	TMD-βL-geph⁻^⁷	0.09/0.27/0.62	0.15/0.44/0.86
Parameter	GlyRβ variant	Synaptic (thresholded)^⁸ Q1/Q2/Q3	Extrasynaptic (thresholded)^⁹ Q1/Q2/Q3
Explored area [10⁻³ µm²]^¹	GlyRβ-wt^¹⁰	0.54/0.73/1.05	0.60/0.91/1.76
	GlyRβ-HI^¹¹	0.55/0.77/1.18	0.63/0.96/1.88
	GlyRβ-LO^¹²	0.58/0.83/1.40	0.77/1.47/3.04
	GlyRβ-D407P/F408G^¹³	0.60/0.86/1.52	0.78/1.52/3.13
	GlyRβ-geph⁻^¹⁴	0.84/1.51/2.73	0.92/1.83/3.48
D_eff [µm² * s⁻¹]^²	GlyRβ-wt¹⁰	0.03/0.04/0.06	0.03/0.05/0.11
	GlyRβ-HI^¹¹	0.03/0.04/0.07	0.03/0.05/0.11
	GlyRβ-LO^¹²	0.03/0.04/0.09	0.04/0.09/0.19
	GlyRβ-D407P/F408G^¹³	0.03/0.04/0.09	0.04/0.09/0.20
	GlyRβ-geph⁻^¹⁴	0.04/0.09/0.19	0.05/0.11/0.22

Median values (Q2), 25% (Q1), and 75% (Q3) quartiles of the explored trajectory areas and D_eff from >700 synaptic trajectories and >7000 extrasynaptic trajectories for each construct.
↵ ¹Explored area (in 10⁻³ µm²).
↵ ²D_eff (in µm²/s).
↵ ³GlyR β-loop (residues 334–455) C-terminally fused to TMD and Dendra2.
↵ ⁴GlyR β-loop with deletion Δ378–393.
↵ ⁵GlyR β-loop with deletion Δ397–410.
↵ ⁶GlyR β-loop with substitution D407P and F408G.
↵ ⁷GlyR β-loop with substitution F398A and I400A.
↵ ⁸Values for synaptic trajectories.
↵ ⁹Values for extrasynaptic trajectories.
↵ ¹⁰mEos4b-tagged full-length human GlyRβ subunit.
↵ ¹¹Full-length GlyRβ with deletion Δ378–393.
↵ ¹²Full-length GlyRβ with deletion Δ414–426].
↵ ¹³Full-length GlyRβ with substitution D407P and F408G.
↵ ¹⁴Full-length GlyRβ with substitution F398A and I400A. For the full-length GlyRβ variants, a threshold of 0.02 µm²/s was applied (corresponding to the median D_eff of GlyRβ-wt in fixed samples). The given Q1, Q2 and Q3 values only represent the trajectories above this threshold.