Figure 3.

Inhibition of cleavage by E165Q mutation in the HEWAHL amino acid motif of CLCA-N domain. Immunoblot analysis of the wildtype and E165Q mutant forms of Flag-tagged CLCA2 in cell lysates, immunoprecipitates, and membrane extracts from transfected HEK293T cells. Membrane pellets were denatured in SDS sample buffer and loaded directly. Mock refers to cells transfected with the empty pLex vector. Both IP and the blot were with anti-Flag antibody. The band at 50 kDa represents the primary antibody used for IP. WT denotes wildtype CLCA2. EQ denotes the E165Q mutant. Experiment was repeated at least three times.