CACNA2D2 mRNA and protein expression.

Left panel: the mRNA content of the proband was evaluated by Real Time-PCR and normalized on that of control individuals (n = 4). CACNA2D2 mRNA resulted strongly reduced in the proband (II:1, 18%), father (I:1, 28%) and mother (I:2, 54%) compared to controls (Anova test, p<0.001). Data are shown as mean ± standard error of three independent experiments. Right panel: the protein content was evaluated by Western blot and normalized on that of control individuals (n = 3), using GAPDH as reference protein and loading control. Graph shows data obtained by densitometric analysis. The proband (II:1) showed a 3% of protein expression compared to controls (Anova on Ranks test, p = 0.034), whereas both parents (I:1 and I:2) had ∼50% of expression. Data are expressed as mean ± standard error of three independent experiments. Representative western blot is shown.