Fig. 5. knockdown of NUP50 in mouse neuronal cells.

a Representative images of western blots for NUP50, different nucleoporins and TDP-43. Western blots were processed in parallel to avoid cross reaction of similar-sized proteins, and quantification of the studied protein was normalized with StainFree loading control for each gel, as provided in Source Data. n = 3 biologically independent experiments, each performed in duplicate. b Dot plots showing a decrease in NUP50 levels (Two-tailed Nested t test: t = 8124, df = 14, ****p < 0.0001) but not other associated nucleoporins, RanGAP or TDP-43 (p > 0.05) after knock-down of the Nup50 mRNA (si-Nup50) compared to the control condition. c, d Representative images and dot plots showing an increase in cytoplasmic inclusions of nucleoporins as stained with mAb414 recognizing the repeated FXFG repeat sequence in nucleoporins in HT22 cell line upon Nup50 knock-down (Two-tailed Nested t test: t = 6,778, df = 16,****p < 0.0001). e, f Representative images and dot plots showing an increase in p62 inclusion in HT22 cell line upon Nup50 knock-down (Two-tailed Nested t test: t = 9 846, df = 17,****p < 0.0001). g, h Dot-plots showing a significant increase in neuronal death (g) in HT22 cell lines (Two-tailed Nested t test: t = 3721, df = 24, **p = 0.0011) and in mouse primary neurons (h) (Two-tailed Nested t test: t = 3,18, df = 34, **p = 0.0031) For all panels, data are presented as mean values ± SEM. n = 3 independent experiments performed at least in duplicate. Each dot in the scatter plot indicates the mean of an individual experiment. All experiments were performed 24 h after transfection.