Journal: Translational Vision Science & Technology
Article Title: Pathogenicity and Functional Analysis of Multi-Variant Allele of RPE 65 Causing Retinitis Pigmentosa
doi: 10.1167/tvst.15.2.1
Figure Lengend Snippet: Pathogenic variant damaged the stability of RPE65 possibly through the ubiquitination–proteasome pathway. ( A ) When HEK293T cells were transfected with WT variant plasmids for 24 hours and treated with 100-µM CHX, RPE65 expression was detected. ( B ) RPE65 stability was detected after cells were treated with 30-µM MG-132. ( C ) Twenty-four hours after transfection, WT mutant proteins were enriched by immunoprecipitation, and their ubiquitination levels were detected.
Article Snippet: After 24 hours of transfection, HEK293T cells were treated with 100-μM cycloheximide (CHX, HY-N0901; MedChemExpress, Monmouth Junction, NJ) or CHX mixed with MG-132 proteasome inhibitor (HY-12320; MedChemExpress).
Techniques: Variant Assay, Ubiquitin Proteomics, Transfection, Expressing, Mutagenesis, Immunoprecipitation