Additional file 2: Figure S1. of Pathogenic LRRK2 variants are gain-of-function mutations that enhance LRRK2-mediated repression of β-catenin signaling

Effects of loss of Lrrk2 on tibial cortical Cross Sectional Area, Mean Cortical Thickness, and Ellipticity. Images in Figures A, B and C show i) values for cross-sectional area, mean cortical thickness and ellipticity, respectively in female wild-type (WT) and Lrrk2 knockout (KO) mice; together with ii) the same data expressed as a graphical heat map along the tibial length; and iii) the points at which differences between genotypes for these parameters become statistically significant. (blue = n/s, yellow, p < 0.05, green p < 0.01, red, p < 0.001). (PDF 131 kb

Additional file 3: Figure S2. of Pathogenic LRRK2 variants are gain-of-function mutations that enhance LRRK2-mediated repression of β-catenin signaling

Effects of loss of Lrrk2 on tibial cortical Zmax and Zmin. Images in Figures A and B show i) values for Zmax and Zmin, predicted resistance to fracture along the shortest and longest cross-sectional axes, respectively in female wild-type (WT) and Lrrk2 knockout (KO) mice; together with ii) the same data expressed as a graphical heat map along the tibial length; and iii) the points at which differences between genotypes for these parameters become statistically significant. (blue = n/s, yellow, p < 0.05, green p < 0.01, red, p < 0.001). (PDF 110 kb

Spectral dynamics of polarization-rotating vector solitons

Vector soliton is obtained by using a fiber stretcher inside a dispersion-engineered nonlinear-polarization-rotation (NPR) mode-locked fiber laser. Fascinating real-time spectral dynamics of vector soliton is observed for the first time using dispersive Fourier transform (DFT).link_to_subscribed_fulltex

Additional file 6: Figure S5. of Pathogenic LRRK2 variants are gain-of-function mutations that enhance LRRK2-mediated repression of β-catenin signaling

Co-localisation between mycLRRK2 and HA-AXIN1, and mycLRRK2 and HA-GSK3β in HEK293 cells. A) Recruitment of mycLRRK2 (green) into polymers formed by HA-AXIN1 (red). Magnified images of a selected region are included. B) shows cytoplasmic co-localisation between mycLRRK2 (green) and HA-GSK3β (red), also with magnified images included. Note that in both experiments, counterstaining with phalloidin (blue) and DAPI (grey) were performed to visualise filamentous actin and chromosomal DNA respectively. The scale bar = 10 μm. (PDF 92 kb

Emulating Reverb and Delay Units Using Fast Convolution

Architecture & Allied Art

Additional file 7: Figure S6. of Pathogenic LRRK2 variants are gain-of-function mutations that enhance LRRK2-mediated repression of β-catenin signaling

LRRK2 interacts with β-catenin and represses β-catenin-driven TOPflash activity. A) HEK293 cells were co-transfected with FLAG-tagged β-catenin (lanes 1 and 4), myc-tagged LRRK2 (lanes 2 and 5), or myc-tagged LRRK2 and FLAG-tagged β-catenin (lanes 3 and 6) for 24 h prior to lysis. Lysates were immunoprecipitated with anti-FLAG antibodies and bound protein was resolved by Western blot (lanes 4–6), with the original lysates run alongside to confirm expression of transfected protein (lanes 1–3). Myc-tagged LRRK2 can clearly be seen in anti-FLAG immunoprecipitates from co-transfected cells (lane 6, upper panel). Full images of all blots are also shown. B) HEK293 cells were transfected with the TOPflash or FOPflash reporter plasmids with the indicated combinations of mycLRRK2, FLAG-β-catenin or appropriate control vectors. After 24 h lysates were taken and luciferase activity measured. 1-way ANOVA of TOPflash values revealed a significant effect of transfection on canonical Wnt activity (n = 9, F = 44.893, p < 0.001). Post-hoc Bonferroni testing showed significantly increased Wnt signalling caused by FLAG-β-catenin transfection in the presence or absence of mycLRRK2 co-transfection (p < 0.001 in both cases). Importantly however, co-transfected mycLRRK2 significantly weakened the TOPflash activation elicited by FLAG-β-catenin (p < 0.001). ANOVA of control FOPflash values also revealed significant effects of treatment, however Bonferroni post-hoc analysis found no significant differences in any pair-wise comparison. (PDF 129 kb

Additional file 4: Figure S3. of Pathogenic LRRK2 variants are gain-of-function mutations that enhance LRRK2-mediated repression of β-catenin signaling

Increased canonical Wnt activity in Lrrk2 knockout cells. A, B) Cells were transfected with TK-renilla and TOPflash or FOPflash in the presence of A) FLAG-DVl1 or C) GFP-FZD5 and HA-LRP6, or appropriate vector controls. A) 1-way ANOVA (n = 9; F = 146.199, p < 0.001) followed by Bonferroni post-hoc analysis revealed increased DVL1-driven TOPflash activity in Lrrk2 knockout cells (p < 0.001 versus all other conditions). No significant changes in FOPflash values were detected (n = 9; F = 2.668, p = 0.064). B) 1-way ANOVA (n = 9; F = 70.694, p < 0.001) followed by Bonferroni post-hoc analysis revealed increased GFP-FZD5/HA-LRP6-driven TOPflash activity in Lrrk2 knockout cells (p < 0.001 versus all other conditions). By contrast, the same treatment elicited a significant decrease in FOPflash values (1-way ANOVA: n = 6; F = 11.129, p = 0.001. Bonferroni post-hoc analysis p < 0.001). C) Wild-type and Lrrk2 MEFs were co-transfected with TK-renilla and TOPflash or FOPflash in the presence of active or inactive (KR) HA-tagged CK1ε. 1-way ANOVA (n = 9; F = 7.619, p = 0.001) followed by Bonferroni post-hoc analysis revealed increased CK1ε-driven TOPflash activity in Lrrk2 knockout cells (p < 0.01 versus all other conditions). No significant changes in FOPflash values were detected (n = 3; F = 1.535, p = 0.279). (PDF 90 kb

Downregulated Wnt/β-catenin signalling in the Down syndrome hippocampus.

Pathological mechanisms underlying Down syndrome (DS)/Trisomy 21, including dysregulation of essential signalling processes remain poorly understood. Combining bioinformatics with RNA and protein analysis, we identified downregulation of the Wnt/β-catenin pathway in the hippocampus of adult DS individuals with Alzheimer's disease and the 'Tc1' DS mouse model. Providing a potential underlying molecular pathway, we demonstrate that the chromosome 21 kinase DYRK1A regulates Wnt signalling via a novel bimodal mechanism. Under basal conditions, DYRK1A is a negative regulator of Wnt/β-catenin. Following pathway activation, however, DYRK1A exerts the opposite effect, increasing signalling activity. In summary, we identified downregulation of hippocampal Wnt/β-catenin signalling in DS, possibly mediated by a dose dependent effect of the chromosome 21-encoded kinase DYRK1A. Overall, we propose that dosage imbalance of the Hsa21 gene DYRK1A affects downstream Wnt target genes. Therefore, modulation of Wnt signalling may open unexplored avenues for DS and Alzheimer's disease treatment