Fig. 4

RIG-I and MDA5 agonists stimulate the expression of IFN-β in hNSCs. a, b Human NSCs were transfected with 5′pppRNA or poly(I:C) HMW. Total RNA was collected at 12 and 24 h post-transfection, and RT-qPCR assay was performed to examine the relative amount of IFN-β mRNA (a). The expression of phospho-IRF3 and total IRF3 was detected through a western blot assay. β-actin was used as an internal control. (b). c Human NSCs were transfected with siRNA specific for RIG-I or MDA5 by Lipofectamine RNAiMAX 2000 and then transfected with 1 μg 5′pppRNA for 24 h. RT-qPCR analysis was performed to detect the expression of IFN-β transcripts while western blot was applied to confirm the knockdown efficiency. d Human NSCs were transfected with siRNA targeting RIG-I or MDA5 and then transfected with 1 μg of poly(I:C) HMW for 24 h. RT-qPCR analysis was performed to detect the expression of IFN-β transcripts while western blot was applied to confirm the knockdown efficiency. The experiments were performed in triplicate, and the error bars represented the SD. Student’s t test was used for statistical analysis. *p < 0.05, **p < 0.01, ***p < 0.001