Fig. 5: The LRRK2-P1446L mutation recapitulates DAPK1/NF-κB activation and apoptosis in vitro.

Representative blots and quantification of (a, b, d, e) LRRK2, Bcl-2, and Bax levels in WT and LRRK2-P1446L MN9D cells and (c, f) DAPK1, p-IκBα, IκBα, p-AKT, AKT, p-NF-κB, and NF-κB in WT and LRRK2-P1446L BV2 cells; n = 3 per group. The blots shown in Fig. 5 were cropped for clarity. The corresponding uncropped scans are presented in Fig. S9 - Part V and VI. g, h Immunostaining and quantification of DAPK1 intensity in TH⁺ MN9D cells. Scale bar, 40 μm (main); 8 μm (inset). The magnified insets are shown on the right. n = 6 per group. i, j MitoSOX fluorescence and quantification in MN9D cells. Scale bar, 20 μm (main); 5 μm (inset). The magnified insets are shown on the right. n = 6 per group. k, l TUNEL staining and quantification of apoptosis in MN9D cells. Scale bar, 50 μm (main); 10 μm (inset). n = 6 per group. The data are presented as the means ± SEMs. For d–f, significance was determined using an unpaired two-tailed Student’s t test (*p < 0.05 and **p < 0.01). For h, j and l, significance was determined by one-way ANOVA with Tukey’s post hoc test (**p < 0.01 vs. Vector group; ##p < 0.01 vs. WT group).