Fig. 4.

Role of CREB in RNS60-mediated increase in myelin genes in primary mouse oligodendrocytes. After different minutes of stimulation with 10% v/v RNS60 or NS under serum free conditions, cell lysates were prepared and analyzed by Western blotting with antibodies against phospho-CREB and total CREB (a). Bands were scanned and values (P-CREB/CREB) are presented as relative to control (b). Results are means ± SD of three different experiments. ^a p < 0.001 vs. control. After different minutes of stimulation with 10% v/v RNS60 under serum free conditions, nuclear extracts were examined for EMSA (c). Results represent three independent experiments. Cells were transfected with CREB siRNA and control siRNA using Lipofectamine Plus and nupherin reagent. After 24 h of transfection, cells were treated with 10% v/v RNS60 under serum-free condition for 6 h followed by monitoring the mRNA expression of MBP and MOG by semi-quantitative RT-PCR (d) and real-time-PCR (e). Results are mean ± SD of three separate experiments. ^a p < 0.001 vs. control; ^b p < 0.001 vs. control-RNS60