Figure 1. LRRK2 Overexpression Enhances Fur1 Translation in DA Neurons.

(A) Dorsal view of a whole-mount 10-day-old female Drosophila brain (yw) stained with anti-Fur1 (green).

(B–D) Left: adult Drosophila brain stained with anti-TH (red) (B), anti-Fur1 (green) (C), and merged (D). Right: magnification of the PPL1 cluster in boxed region for (B)–(D).

(A-D) Imaging was performed at 63x magnification. Thirty-six images were captured and tiled following a rectangular grid mode using Zen software.

(E–G) PPL1 clusters double stained with anti-TH (red) and anti-Fur1 (green) in flies expressing (E) UAS-LacZ (Ddc-Gal4/UAS-LacZ), (F) UAS-LRRK2^IT (Ddc-Gal4/+; UAS-LRRK2^IT/+) and (G) UAS-S6K^STDE (Ddc-Gal4/+; UAS-S6K^STDE/+) in dopaminergic (DA) neurons.

(H) Quantification of Fur1 sensor fluorescence intensity relative to TH for genotypes in (E)–(G). Each data point represents fluorescence from a single neuron. n = 100 neurons from 10 PPL1 clusters for each genotype. Data are represented as mean ± SEM. One-way ANOVA with Bonferroni post-test.

(I–L) PPM1-2 clusters expressing Fur1 sensor (5′UTR of Fur1 upstream of EGFP) (green) and stained with anti-TH (red) in flies co-expressing (I) UAS-LacZ (UAS-Fur1-sensor/UAS-LacZ; Ddc-Gal4/+), (J) UAS-LRRK2^IT (UAS-Fur1-sensor/+; Ddc-Gal4/UAS-LRRK2^IT), (K) UAS-S6K^STDE (UAS-Fur1-sensor/+; Ddc-Gal4/UAS-S6K^STDE) and (L) UAS-4E-BP^AA (UAS-Fur1-sensor/+; Ddc-Gal4/UAS-4E-BP^AA) in DA neurons. GFP corresponding to Fur1 sensor (green) is directly imaged without antibody staining.

(M) Quantification of Fur1 sensor fluorescence intensity relative to TH for genotypes in (I)-–(L). Each data point represents fluorescence from a single neuron. n = 50 neurons from 10 PPM1-2 clusters for each genotype. Data are represented as mean ± SEM. One-way ANOVA with Bonferroni post-test.

*p < 0.05, **p < 0.01, ***p < 0.001.