Figure 1.

AP1S3 loss-of-function mutations are most likely to affect skin keratinocytes. (a) Schematic representation of AP-1 structure. (b) HEK293 cells were transfected with wild-type and mutant AP1S3 constructs. Lysates were subjected to the indicated temperature gradient, and soluble (nondenatured) proteins were analyzed by Western blotting. The densitometry shows the fraction of nondenatured protein (mean ± standard error of the mean of the results obtained in two experiments). (c) HEK293 cells were transfected with myc-tagged AP1S3 and FLAG-tagged AP1M1 constructs. Lysates were subjected to immune precipitation (IP) and immune blotting (IB) as indicated. The image is representative of results obtained in two experiments. (d) Real-time PCR analysis showing abundant AP1S3 expression in keratinocytes. The data show the mean ± standard error of the mean of measurements obtained in two donors. ^∗P ≤ 0.05. IB, immune blotting; IP, immune precipitation; WCE, whole cell extracts; wt, wild type.