Co-immunoprecipitation of FLAG-WT ADA2 and HA-ADA2 G47R, R169Q, T360A, or H424N supports interaction between WT ADA2 and mutant ADA2. (A) Illustration depicting experimental workflow used for B and C. (B) Representative confocal microscopy image of cotransfected HEK293T cells stained for FLAG-tagged WT ADA2, HA-tagged MT ADA2, and nucleus illustrating successful cotransfection of differentially tagged plasmids. Images were acquired on Andor Dragonfly Confocal Spinning Disk at 63× magnification. (C) Whole-cell lysates of HEK293T cells transiently cotransfected with FLAG-WT ADA2 and HA-ADA2 G47R, R169Q, T360A, or H4242 were co-immunoprecipitated with an anti-FLAG antibody. Western blot analysis of the whole-cell lysate and eluate of immunoprecipitation with anti-HA, anti-FLAG, anti-ADA2, and anti-B-actin. The image shown represents two independent experiments. EV, empty vector; IB, immunoblot. Source data are available for this figure: SourceData FS3.
Figure S3.

Co-immunoprecipitation of FLAG-WT ADA2 and HA-ADA2 G47R, R169Q, T360A, or H424N supports interaction between WT ADA2 and mutant ADA2. (A) Illustration depicting experimental workflow used for B and C. (B) Representative confocal microscopy image of cotransfected HEK293T cells stained for FLAG-tagged WT ADA2, HA-tagged MT ADA2, and nucleus illustrating successful cotransfection of differentially tagged plasmids. Images were acquired on Andor Dragonfly Confocal Spinning Disk at 63× magnification. (C) Whole-cell lysates of HEK293T cells transiently cotransfected with FLAG-WT ADA2 and HA-ADA2 G47R, R169Q, T360A, or H4242 were co-immunoprecipitated with an anti-FLAG antibody. Western blot analysis of the whole-cell lysate and eluate of immunoprecipitation with anti-HA, anti-FLAG, anti-ADA2, and anti-B-actin. The image shown represents two independent experiments. EV, empty vector; IB, immunoblot. Source data are available for this figure: SourceData FS3.

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