Fig. 5: Knockdown of spinal Fbxo45 expression impedes Fbxo45-dependent Munc13-1 ubiquitination to induce behavioral allodynia. | Cell Death & Disease

Fig. 5: Knockdown of spinal Fbxo45 expression impedes Fbxo45-dependent Munc13-1 ubiquitination to induce behavioral allodynia.

From: Spinal TNF-α impedes Fbxo45-dependent Munc13-1 ubiquitination to mediate neuropathic allodynia in rats

Fig. 5

a Representative Western blot and statistical analysis (normalized to GAPDH) demonstrating that intrathecal administration of Fbxo45 siRNA (Fbxo45 siRNA; 1, 3, and 5 μg; 10 μL; once daily for 4 days), but not missense siRNA (MS siRNA, 3 μg, 10 μL) or polyethylenimine (a transfection reagent, PEI, 10 μL), dose-dependently decreased Fbxo45 expression in the dorsal horn of naive rat. it, implantation of an intrathecal catheter. IB immunoblotting. **P < 0.01 compared with naive (n = 6). b Intrathecal application of neither Fbxo45 siRNA (Fbxo45 siRNAi, 3 μg, 10 μL) nor missense siRNA (MS siRNA, 3 μg, 10 μL) to naive rats resulted in motor deficits (Rotarod test). The gray bar at the bottom indicates the duration of intrathecal administration (n = 7). c Intrathecal administration naive rats with Fbxo45 siRNA (Fbxo45 siRNA; 3 μg, 10 μL), decreased the withdrawal threshold of the hind-paw at days 2, 3, and 4 after the start of injection (von Frey test). The gray bar at the bottom indicates the duration of administration. *P < 0.05, **P < 0.01 compared with naive (n = 7). d The intrathecal application of Fbxo45 siRNA (Fbxo45 siRNAi, 3 μg, 10 μL) into naive rats decreased Munc13-1 ubiquitination in the synaptic plasma membranes of the ipsilateral dorsal horn. SPM IP synaptic plasma membranes immunoprecipitation. Sham 7D, sham operation at day 7. **P < 0.01 compared with naive (n = 5). e Representative Western blot and statistical analysis (normalized to N-Cad) demonstrating that intrathecal administration naive rats with Munc13-1 siRNA (Munc13-1 siRNA; 3 μg, 10 μL; once daily for 4 days) decreased the Munc13-1 expression without affecting the Fbxo45 expression in the SPM of the ipsilateral dorsal horn. Intrathecal administration naive rats with Fbxo45 siRNA (Fbxo45 siRNA; 3 μg, 10 μL; once daily for 4 days) increased the Munc13-1 expression, but decreased the Fbxo45 expression in the SPM of the ipsilateral dorsal horn. **P < 0.01 compared with naive (n = 6). Results were expressed as mean ± SEM

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