Journal: bioRxiv

Article Title: The Parkinson’s disease associated Leucine-rich repeat kinase 2 affects expression of Transferrin receptor 1 and phosphorylation of key signaling proteins in human iPSC-derived dopaminergic neurons

doi: 10.1101/2025.05.06.652393

Figure Lengend Snippet: (a) Western blot analysis and quantification of TfR protein expression in hG2019S-transgenic (hG2019S tg), Lrrk2-deficient (Lrrk2 k.o.) and wild-type fibroblasts (n = 8 independent experiments). (b) Transferrin recycling assay in rat fibroblasts: Phalloidin (green), DAPI (data not shown) and TexasRed transferrin (white) staining were used for automated quantification of generated data (segmentation and ROI count). Representative images for wild-type fibroblasts, time points 0 und 70 min (top). Graphical representation of transferrin uptake at different time points (= incubation time) in hG2019S tg (left, n = 11 independent experiments) and Lrrk2 k.o. (right, n = 9 independent experiments) fibroblasts (button). (c) TfR protein degradation at different time points after cycloheximide addition (n = 8 independent experiments). (d) Western blot analysis/ quantification of Rab12 expression in hG2019S tg, Lrrk2 k.o. and wild-type fibroblasts.

Article Snippet: To investigate the impact of TfR downregulation on its internalization, we observed the TfR translocation by Transferrin Recycling Assay using the TexasRed-labeled transferrin and BD Pathway 855 imaging system in the same rat cell lines.

Techniques: Western Blot, Expressing, Transgenic Assay, Staining, Generated, Incubation