There is increasing evidence that autophagy contributes to the epidermal differentiation; however, the role of autophagy in epidermal tight junction (TJ) barrier remains unclear. To evaluate the role of autophagy in the maintenance of skin TJ barrier, we knocked out autophagy in human primary keratinocytes by infecting cells with autophagy-related gene 3 (Atg3) C264S mutant adenovirus. Overall design: A total of 4 samples were analyzed and data from autophagy-deficient keratinocytes and normal keratinocytes were compared.