Fig 2: eRNA E2-3 selectively regulates HO-1 induction. (A) The effect of eRNA E2-3-KD on eRNA E2-3 expression. HeLa cells were transfected with control siRNA (Ctrl) or two siRNAs against eRNA E2-3 (E2-3(1) or E2-3(2)) and were untreated (-) or exposed to 100 µM DEM for 6 h (+). The eRNA E2-3 level was analyzed by real-time RT-PCR using specific primers and Universal Probe Library Probe. (B and C) The effect of eRNA E2-3-KD on the expression of HO-1 (B) and TXNRD1 (C). The expression of HO-1 and TXNRD1 was analyzed by real-time RT-PCR using the TaqMan® Gene Expression Assay system. Each value was normalized to cyclophilin A gene expression, and the arbitrary RNA level was expressed as the mean ± SEM of four independent assays. **P < 0.01 (two-tailed unpaired Student's t-test) compared to control siRNA without DEM (Ctrl (-)). #: P < 0.05; ##: P < 0.01; NS, not significant compared to the value of control siRNA with 100 µM DEM for 6 h (Ctrl (+)) (one-way ANOVA followed by a Dunnett's post hoc test for multiple parameter comparisons). (D) The effect of eRNA E2-3-KD on HO-1 protein expression. Whole-cell lysates were separated by SDS-PAGE and HO-1 protein expression was analyzed by immunoblotting using a HO-1-specific antibody (Abcam, ab68477). Actin was used as a loading control (Sigma-Aldrich, A1978).

Fig 3: Western blot detection of HIP1 in cerebellar white matter lysates (N = 29). Immunoblots were labelled with anti-HIP1 (Abcam ab181238, 1:2000), anti-ß-actin (Sigma A1978, 1:5000) and IRDye secondary antibodies (LiCor), and scanned on a LiCor Odyssey Fc (A). Band intensities were derived from raw scan data and analysed on Image Studio (LiCor). HIP1 band intensities normalized against ß-actin as housekeeping gene (B). *Horizontal black bars indicate significant differences (adjusted p < 0.05 using Wilcoxon rank-sum test with Benjamini–Hochberg adjustment for multiple testing). CTRL, healthy controls; HD, Huntington's disease; MSA, multiple system atrophy; PD, Parkinson's disease; PSP, progressive supranuclear palsy