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. 2015 Jul 21;290(36):21939–21950. doi: 10.1074/jbc.M115.657668

FIGURE 5.

FIGURE 5.

A GVVASQPARV sequence of SNAP25b is sufficient for specific recognition by the AR domain of zDHHC17/13. A, HA-tagged AR-containing fragments of zDHHC17 and zDHHC13 (STOP codons on cDNAs introduced at corresponding amino acid positions marked with X) were expressed in HEK293T cells, and their ability to interact with the GVVASQPARV peptide (SNAP25b111–120) and full-length SNAP25b (SNAP25bFL) was assessed by pulldowns of corresponding lysates by GST, GST-SNAP25b111–120, and GST-SNAP25bFL. One-twelfth of total inputs and one-quarter of total bound fractions were run on 12% gels, and following transfer, blots were stained by Ponceau S solution and probed with an HA antibody. B, HEK293T cells expressing HA-zD17301X, HA-zD13290X, or FLAG-TANK2 were lysed, and the binding preference of these AR proteins to a ΨβXXQP (SNAP25b peptide) or a RXXDPG (IRAP peptide) sequence was assessed by pulldowns of corresponding lysates by GST, GST-SNAP25b111–120, and GST-IRAP78–109. Bound fractions and one-twelfth of total inputs were run on 12% gels, and following transfer, blots were stained by Ponceau S solution and probed with either HA or FLAG antibodies.