Fig. 9: Impaired degradation of ISGylated Atxn1 protein in human Purkinje cells. | Communications Biology

Fig. 9: Impaired degradation of ISGylated Atxn1 protein in human Purkinje cells.

From: Dynamic molecular network analysis of iPSC-Purkinje cells differentiation delineates roles of ISG15 in SCA1 at the earliest stage

Fig. 9

a ISGylated proteins were immunoprecipitated with an anti-ISG15 antibody from post-mortem cerebellar tissues of normal control and SCA1 patients and blotted with an anti-ISG15, anti-Atxn1, or anti-Ub antibody. The images were the result with a representative patient (28/53 CAG repeats). b Quantitative analysis of the signal intensities of Atxn1 or ubiquitinated Atxn1 bands in immunoprecipitates obtained using an anti-ISG15 antibody. N = 3 (see Methods). c Chase assay of Atxn1 protein degradation after inhibition of protein translation by cycloheximide in human iPSC-derived Purkinje cells. Degradation of mutant Atxn1 was delayed in comparison with that of normal Atxn1 and was normalized by siRNA-ISG15. The enhancement of protein degradation by siRNA-ISG15 was blocked by the proteasome inhibitor MG132, but was unaffected by the autophagy inhibitor bafilomycin A. d Quantitative analysis of the signal intensities of Atxn1 bands during the chase assay. N = 4.

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