Fig. 8: Effects of HsATG4D p.S89N and p.Y280C allelic variants. | Cell Death & Differentiation

Fig. 8: Effects of HsATG4D p.S89N and p.Y280C allelic variants.

From: ATG4D is the main ATG8 delipidating enzyme in mammalian cells and protects against cerebellar neurodegeneration

Fig. 8

A Up, predicted impact of p.S89N and p.Y280C variants using publicly available prediction algorithms. Numbers indicate the probability of a determined variant to be neutral (green boxes) or detrimental (red boxes) for protein structure stability. Middle, schematic representation of the different protein domains present in ATG4D. Bottom, scheme showing conservation of S89 and Y280 residues across evolutionarily-distant vertebrate species. B Computer-based model of HsATG4D 3D structure interacting with a substrate and an auxiliary LC3B. The position of p.S89N and p.Y280C variants and adjacent residues are shown in more detail. C Immunoblotting analyses against mATG8 proteins in WT, Atg4d−/− MEFs and Atg4d−/− MEFs stably expressing either Flag-HsATG4D, Flag-HsATG4D p.S89N or Flag-HsATG4D p.Y280C in the indicated conditions. D Representative images of immunofluorescence against endogenous mATG8 proteins in WT, Atg4d−/− MEFs and Atg4d−/− MEFs stably expressing either Flag-HsATG4D, Flag-HsATG4D p.S89N or Flag-HsATG4D p.Y280C in the indicated conditions. Images show cells cultured in full medium. E Quantification of the data from (D). LOAD: β-actin. Bars represent mean ± SD (N > 80 cells per condition). Scale bar: 10 μm. *P < 0.05, 2-tailed unpaired Student’s t test.

Back to article page