Fig. 1
Expression of the GABAergic system and cell viability assay in OUMS-27 cells. a Determination of the mRNA levels of GAD65, GAD67, the GABAA α1–6, β1–3, γ1–3, δ, π, θ, and ε subunits, and GABAB R1a, R1a/b, and R2 in OUMS-27 cells by RT-PCR. b Confocal microscopy of the GABA, GAD, GABAA receptor subunits, and GABAB receptor subunits in OUMS-27 cells (a- j). (a) GABA, (b) GAD65, (c) GAD 67, (d) goat IgG, (e) α2, (f) α3, (g) β1, (h) γ3 (i) R1, and (j) R2. Immunoreactivity is visible as green fluorescence and cell nuclei are stained with PI (red). Arrow heads indicate immunoreactive cells. Scale bar = 10 μm. c Cell viability assay; OUMS-27 cells were treated with 100 μM GABA, 50 μM MUS (GABAA receptor agonist), 100 μM BFN and 10 μM SKF (GABAB receptor agonists), 100 μM GABA+ 100 μM BMC (GABAA receptor antagonist) or 100 μM GABA+ 1 μM CGP (GABAB receptor antagonist). The cell proliferation ELISA and BrdU assays were performed after drug treatment. Colorimetric analysis was performed using an ELISA plate reader. ** indicates significant differences between the control and each group (P < 0.01). Data are presented as the mean ± SD