Fig. 2.

TDP-43-associated cryptic splicing across phosphorylated TDP-43 pathological subtypes of ALS. a–b Expression levels of UNC13A cryptic RNA a and truncated STMN2 RNA b measured by quantitative real-time PCR across ALS pathological subtypes (Type 1, n = 12; Type 2a, n = 12; Type 2b, n = 11; total n = 35). UNC13A cryptic RNA and truncated STMN2 RNA levels were normalized to their respective full-length transcripts using the ΔCt method. Statistical comparisons were performed using ΔCt values with the Kruskal–Wallis test followed by Dunn’s multiple-comparisons test. Data are shown as individual values; horizontal bold lines indicate the median, and error bars represent the interquartile range (25th–75th percentiles). Asterisks indicate significant differences (*p < 0.05, **p < 0.01, ***p < 0.001); ns indicates not significant. c Correlation between UNC13A cryptic RNA and truncated STMN2 RNA expression levels across ALS cases. Expression levels were normalized to the corresponding full-length transcripts using the ΔCt method. Correlation was assessed using Spearman’s rank correlation coefficient. Each point represents an individual case. The Spearman correlation coefficient (ρ) and 95% confidence interval (CI) are shown in the figure. Asterisks indicate significant correlations (***p < 0.001). An inset provides a zoomed view of the low-expression range (UNC13A cryptic RNA: 0–0.005; truncated STMN2 RNA: 0–0.02)