Fig. 4: FAT10 stabilises NCOA4 expression by inhibiting its ubiquitination in pancreatic acinar cells. | Cell Death & Disease

Fig. 4: FAT10 stabilises NCOA4 expression by inhibiting its ubiquitination in pancreatic acinar cells.

From: Mechanistic insights into the role of FAT10 in modulating NCOA4-mediated ferroptosis in pancreatic acinar cells during acute pancreatitis

Fig. 4

A AR42J cells were treated with MG132 (15 μM) for the indicated time, NCOA4 levels were detected by western blotting; B Quantitive shotgun analysis showed substrate proteins interaction with FAT10; C Co-IP for FAT10 and NCOA4 in AR42J cells; D GST-pull-down analysis for FAT10 and NCOA4 in AR42J cells; E Colocalisation of FAT10 and NCOA4 in AR42J cells (Scale bar: 10 μm); F Western blotting analysis of FAT10 and NCOA4 expression in AR42J cells transfected with the indicated plasmids treated with or without MG132 (15 μM); G The cells were exposed to CHX (20 μM) at the indicated times, and the degradation of exogenous FAT10 and NCOA4 was detected in AR42J cells transfected with the indicated plasmids; H AR42J cells were transfected with increasing amounts of Flag-FAT10 plasmid. The cells were lysed for immunoprecipitation using anti-Ub and anti-FAT10 beads to detect NCOA4 binding; I Binding of NCOA4 during the course of the competition was analysed by GST-pull-down experiments; J, K AR42J cells transfected with the indicated plasmid were treated with MG132 (15 μM) and the ubiquitination of NCOA4 was detected by western blotting analysis. ***p < 0.001.

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