Figure 6.
SNL-induced overexpression of CaV1.2. Efficient knockdown after intrathecal anti-CaV1.2 antisenses application. A, Expression of CaV1.2 mRNA (qRT-PCR) increased by more than twofold in SNL rats compared to sham animals (black bar). CaV1.2 mRNA levels were significantly reduced after intrathecal injection of anti-CaV1.2 TP10-PNA (white bar) or siRNA (dotted bar). Application of either anti-CaV1.3 TP10-PNA (gray bar) or mismatch PNA or mismatch siRNA (middle and right hatched bars, respectively) had no effect. B, Expression of CaV1.3 mRNA (qRT-PCR) showed a significant reduction in SNL rats compared to sham animals (black bar), and was further decreased after intrathecal injection of anti-CaV1.3 TP10-PNA (gray bar). Application of either anti-CaV1.2 TP10-PNA (white bar) or mismatch PNA (hatched bar) had no effect. C, Western blot analysis of the CaV1.2 expression from ipsilateral lumbar spinal cord of sham, SNL, siRNA-injected, and PNA-injected animals (C1). The protein level increased in the spinal cord of SNL animals (black bar). Application of anti-CaV1.2 PNA or siRNA (white and dotted bars, respectively) suppressed CaV1.2 overexpression (C2). n, Number of animals; one star, p < 0.05; two stars, p < 0.01; three stars, p < 0.001; NS, nonsignificant; stars above black bars are statistics versus sham.