not-yet-known not-yet-known not-yet-known unknown Figure 5. MRS5474 inhibited GAT-1 mediated GABA uptake, an action mimicked by an adenosine A3R agonist and antagonized by an A3R antagonist but not by an A1R antagonist . In both A and B are shown the absolute values (fmol/mg protein) of GAT-1 mediated GABA uptake in control conditions (no drug, ●) or in the presence of MRS5474 (50 nM) (A , ▲) or MRS5698 (100 nM) (A3R agonist, B, ▼ ). Data from the same experiment is connected by a straight line. In C is shown the pooled data as (fmol/mg protein) in: (from left to right) control conditions, in the presence of MRS5474 (50 nM), in the presence of MRS5698 (100 nM). In D is shown the pooled data as % change in GAT-1 mediated GABA uptake caused by: (from left to right) MRS5474 (50 nM), MRS5474 (50 nM) in the presence of MRS1523 (10 µM, an A3R antagonist), MRS5474 (50 nM) in the presence of DPCPX (50 nM, an A1R antagonist); MRS5698 (100 nM). In E and F is shown the pooled data in control conditions (no drug, left column in each panel) or in the presence of MRS1523 (10 µM) (E , right column), or DPCPX (50nM) (F , right column). In panels C-F data is shown as mean ± SEM with the dots representing individual data points in each experiment; *P<0.05; **P<0.01; ***P<0.001; ns: P>0.05, two tailed paired t test (A and B ), or one-way ANOVA with multiple comparisons tests (C : Tukey’s, to compare each column with all other columns; D : Dunnet’s, to compare the 1st column with all other columns) or two tailed unpaired student’s t test (E and F ). Figure 6. A3R immunoreactivity was enhanced in hippocampal samples from patients with refractory epilepsy . Representative western blot of A3R immunoreactivity in epileptic resected tissue from patients with refractory epilepsy and in post-mortem tissue from control patients (CTRL) are shown in A. GAPDH was used as control. B: Pooled data from densitometric analysis of the 35 kDa band (A3R as indicated by the antibody supplier) corrected for GAPDH (37 kDa) in the same samples. Values were normalized taking as 1 the values obtained in control samples. CTRL and epileptic samples were always analyzed in the same membrane. Data in B is shown as mean ± SEM with the dots representing individual data points (sample details in Table 2). *P<0.05 two tailed unpaired t-test with Welch’s correction for unequal variances.