Fig. 5. Beneficial and adverse effects of ASO–CCG (11 nt) at a molecular level in an FXTAS mouse model.

a, b DE, results based on differential gene expression analysis of the RNA-seq data. adj.P.Val, P value generated using moderated t-statistic and adjusted for multiple testing using Benjamini–Hochberg’s method. a Plot, DE for striatum of control, saline- or ASO–CCG-treated P90CGG mice (N = 4 per group). On the x axis, the log₂ fold change in the expression levels of the 416 genes (one outlier gene, Gm20425, was excluded from analysis), that were significantly changed (adj.P.Val < 0.05) in saline-treated P90CGG mice compared to control mice is plotted. On the y axis, the log₂ fold change in the gene expression levels of the same genes compared between ASO–CCG- and the saline-treated group is plotted. A general restoration of the expression levels towards the levels in control mice was observed in the ASO–CCG-treated group (red line, negative Pearson correlation with r = −0.75 and P < 0.001). Bar graphs, gene ontology analysis of genes extracted from DE for striatal tissue according to the following parameters: average expression value >1 and adj.P.Val ≤ 0.05. Significantly upregulated and downregulated genes were analyzed for the saline vs control (gray) and ASO–CCG vs control groups (blue). Red line, P = 0.01 in Fisher’s exact test with Bonferroni correction for multiple testing. b Analysis of expression changes in genes containing CGG repeats in ASO–CCG-treated P90CGG mice. DE for the cortex of saline- or ASO–CCG-treated P90CGG mice (N = 4 for each group). A global comparison of gene expression between these two groups of mice showed a significant increase in the expression of 22 genes containing at least 6 CGG repeats (red line) compared to all analyzed genes (black line; P < 0.001, two-tailed Mann–Whitney test). Moreover, in the list of genes showing significant expression changes between these two groups (adj.P.Val < 0.05), genes containing ≥ 6 CGG repeats were enriched 13-fold relative to the expected level. c Steady-state level of proteins encoded by genes carrying short CGG repeats in 5′UTRs in the cortex of saline- and ASO–CCG-treated P90CGG mice. N = 4 animals. The CGG repeat length is specified for each gene. d RT-qPCR of immune system-related genes in the cortex of P90CGG mice. N = 4 animals. e Steady-state level of immune system-related toxicity markers in P90CGG mice. Hippocampus: GFAP, CD68, N = 6 animals. Samples were derived from the same experiment and processed in parallel on different gels. Cortex: AIF1, N = 4 animals. c, e Western blot. All blots were cropped. c–e Gray bars, saline-; blue bars, ASO–CCG-treated animals. Graphs present means of indicated N, with the SDs. Two-tailed unpaired Student’s t test, **P < 0.01; ***P < 0.001; P > 0.05, ns, non-significant. a–d Source data are provided as a Source Data file.