Fig. 3: Autophagy markers in VMDA and cortical neurons show distinct phenotypes in PRKN and SNCA mutations.

A Representative images of VMDA neurons immunostained with P62 (Magenta), TH (green) and DAPI (blue) in the absence (left panel) and presence (right panel) of Bafilomycin A1. The number of P62 spots per cell in the different mutation groups of VMDA neurons in the absence (B) or presence (C) of Bafilomycin A1. D) Representative images of cortical neurons immunostained with P62 (Magenta), MAP2 (green) and DAPI (blue) in the absence (left panel) and presence (right panel) of Bafilomycin A1. The number of P62 spots per cell in the different mutation groups of cortical neurons in the absence (E) or presence (F) of Bafilomycin A1. Representative immunoblot (G) and subsequent quantification (H) of P62 protein levels in cortical neurons of different mutation groups. Protein expression is normalised to β-actin. I) Representative images of cortical neurons immunostained with Galectin-3 ((magenta), MAP2 (green) and DAPI (blue). J) Representative images of VMDA neurons immunostained with TFEB (magenta), TH (green) and DAPI (blue). The number (K) and size (L) of Galectin-3 spots in the different mutation groups of cortical neurons M) TFEB intensity per cell in the different mutauion groups of VMDA neurons. The scale bar in all images is 200 µm. Image quantification graphs show estimated marginal mean ± SEM. Each datapoint on the graph represents the mean value for each cell line derived from n = 3 biological replications (each performed in at least duplicate).