Fig 2: Minimal MOG protein reduction was observed across CNS regions following neonatal ICV injection of AAV-CRISPR/sgMOG.Viruses (ssAAV-U6-sgMOG-CBA-NLS-eGFP) were injected at either a low (a 5E10 vg) or high dose (b 20E10 vg) in neonatal mice. After 6 weeks, multiple brain regions and spinal cord were dissected and the amount of the MOG proteins remaining undisrupted in the bulk tissues was quantified using the Protein Simple Peggy Sue. For quantification of each AAV variant within each CNS region: the averages of the MOG signals from AAV-CRISPR/sgLacZ (not shown) were normalized to 100% as baseline (dotted line), relative MOG protein levels are presented as mean ± s.e.m., two-way ANOVA followed by Tukey’s test, from baseline (sgLacZ) #p < 0.05.

Fig 3: NF-κB activation in OLs impairs myelin-associated protein expression. a to d qRT-PCR reveals no differences in gene expression of Mbp (a), Plp (b), Mog (c) and Mobp (d) at 3 and 20 wpi in cortex tissue (n = 3–5). e to h Western blot analysis of cortex tissue indicates significantly decreased protein levels of MBP, MOG and PLP1 at 20 wpi in IKK2-CAPLP−CreERT2 mice, while protein levels at 3 wpi were comparable between control and IKK2-CAPLP−CreERT2 animals (n = 3). i Principal component analysis of isolated O4+ OLs (RNA sequencing) at 3 and 20 wpi shows distinct clustering within control and IKK2-CAPLP−CreERT2 groups (n = 4, two brains with identical genotype were pooled to n = 1). j Transcriptome analysis determined mild gene expression changes between IKK2-CAPLP−CreERT2 and control O4+ OLs in genes related to to the GO class (GO:0,042,552) myelination, myelin assembly and maintenance, regulation of myelination and paranodal junction assembly. Out of all genes depicted in the heatmap only Cst7, Tlr2, Igf1, Eif2ak3, Cxcr4, Lpin1, Egr2, Cd9, Dag1, and Epb41l3 were significantly deregulated at both timepoints. At 3wpi additionally Ifng, Hes 5, Cntn2 and Dlg1 reached significance, at 20 wpi Nrg1, Id4, Tmem98 and S100b were found significantly deregulated in addition. Data are shown as mean ± SEM. Grey bars, control; red bars, IKK2-CAPLP−CreERT2. Statistical analysis: Two-way ANOVA multiple comparison test followed by Bonferroni´s post hoc test, (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001 NS, non-significant (p > 0.05))