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. 2015 Sep 20;23(9):724–733. doi: 10.1089/ars.2015.6265

FIG. 3.

FIG. 3.

DUOXA2 interacts covalently with DUOX2. (A) Immunoprecipitated DUOX2 and mutant DUOX2 C124G were analyzed by SDS-PAGE in nonreducing conditions. Proteins were stained as described under the Materials and Methods section. Bands with a molecular weight superior to 150 kDa were excised from the gel and microsequenced. (B) Covalent interaction between DUOX2 and DUOXA2 was analyzed by Western blot analysis in nonreducing conditions. DUOX2 or mutant DUOX2 C124G was coexpressed with myc-DUOXA2 in HEK293 cells. The expression of DUOX2 was detected by using anti-hDUOX2 and DUOXA2 was detected by using anti-myc. (C) Cells were cotransfected with DUOX2 or mutant DUOX2 and myc-DUOXA2. Anti-myc immunoprecipitates (IP: c-Myc) of the cell lysates were separated by SDS-PAGE in reducing conditions and analyzed by Western blotting for DUOX2 and myc-DUOXA2. Analysis of the cell lysates used as the input in immunoprecipitation is shown in the upper three panels. SDS-PAGE, sodium dodecyl sulfate–polyacrylamide gel electrophoresis.