Figure 5.

Restoration of Cellular Copper Excretion by COMMD1 Gene Supplementation

(A) Representative phase contrast and fluorescence images of wild-type, COMMD1^−/−, and LV-COMMD1-DsRed organoids, showing successful transduction in LV-COMMD1-DsRed organoids

(B) COMMD1 immunoblot of protein extracts from wild-type, COMMD1^−/−, and LV-COMMD1-DsRed organoids, showing successful protein expression (n = 3 biological replicates). Beta-actin was used as the loading control. The COMMD1 protein in LV-COMMD1-DsRed organoids is larger due to the remaining amino acids of the cleavage site P2A added to the vector. Quantification of protein presented as average ± SD.

(C) Copper sensor measurement before and after CuCl₂ treatment of wild-type, COMMD1^−/−, and LV-COMMD1-DsRed organoids (n = 3 biological replicates), showing that LV-COMMD1-DsRed organoids successfully restored copper excretion.

(D) Representative phase contrast images of wild-type, COMMD1^−/−, and LV-COMMD1-DsRed organoids after CuCl₂ treatment. Deterioration of the culture and cell death (arrows) are presented in COMMD1^−/− organoids.

This figure represents results of three repeated experiments with independent COMMD1^−/− organoids.