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. 2019 Nov 9;22:453–465. doi: 10.1016/j.isci.2019.11.013

Figure 1.

Figure 1

Forskolin Reduces GABAAR and Gephyrin Immunoreactivity at Spinal Cord Synapses

(A) Triple immunolabeling of endogenous GlyRs, GABAARs, and gephyrin in cultured rat spinal cord neurons (DIV16). Cells were treated with 20 μM forskolin (For) or kept under control conditions for 30 min (Ctr; vehicle 0.2% ethanol), fixed, and stained with antibodies against the GlyRα1 and GABAARβ3 subunits, and with the rat gephyrin antibody mAb7a (Geph 7a, Synaptic Systems). Scale bar, 5 μm.

(B) Quantification of the integrated fluorescence intensity at synaptic GlyR puncta in the three channels (cell-by-cell analysis, arbitrary units normalized to the control condition in each channel; data are represented as 10%, 25%, 50% (median), 75%, and 90% percentiles; the mean is indicated as a cross; nCtr = 86, nFor = 79 cells from 6 coverslips and 3 independent experiments; ***p < 0.001, ANOVA).

(C) Cumulative probability of the ratio of the GlyR (black traces) or GABAAR fluorescence intensities (red traces) relative to the Geph 7a immunoreactivity at synapses (control: solid lines, nCtr = 1.4 × 104; forskolin: dotted lines, nFor = 1.2 × 104 synapses).