Fig 1: Validation of RNA-seq data by qRT-PCR and Western blot analyses. a The relative mRNA level of DLK and axon guidance genes in infected cells was analyzed by qRT-PCR, normalized to three housekeeping genes and calculated with the ??C T method. The value of mRNA expression for each gene in control cells (pLKO.1) was arbitrarily set to 1. Data are the mean ± SEM (error bars) from three independent experiments carried out in triplicate. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001; ns, p > 0.05. b Representative Western blots showing levels of DLK, Plxna4, Nrp1 and actin in control and DLK-depleted Neuro-2a cells. c Quantitative densitometric measurements of DLK, Plxna4 and Nrp1 protein levels in infected cells. Results are normalized to actin level in control cells, which were set to 1, and represent mean ± SEM (error bars) from three independent experiments. *, p < 0.05; **, p < 0.01; ***, p < 0.001