Figure 4. Detergent solubility assay of FLAG-tagged wild type and mutant CRYGD. Western blotting analysis showed cell type specific solubility changes of CRYGD mutants. When expressed in human lens B3 epithelial cells (upper set of blots), wild type (WT) and all known mutant CRYGD (including R15S), except G165fsX8, were completely soluble in 0.5% Triton X-100 extraction. G165fsX8 was mainly Tx-insoluble. When expressed in COS-7 cells (lower set of blots), R15C and R15S CRYGD became moderately Tx-insoluble, unlike the wild type and P24T and G61C mutants. G165fsX8 remained Tx-insoluble.