Figure 1.

Expression of filaggrin-2 is efficiently inhibited. (a) At day 10, fully differentiated shc- and shFLG2-RHEs were analyzed by qRT-PCR. The expression of the TATA box-binding protein gene was used for normalization. (b) Identical shc- and shFLG2-RHEs were analyzed by western blot with a polyclonal antibody specific for the spacer domain of filaggrin-2 (FLG2). A representative result is shown on the left. Immunodetected filaggrin-2 was quantified and normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) levels (shown on the right). (c) Identical shc- and shFLG2-RHEs were analyzed by immunofluorescence with the same antibody. In shc-RHE, filaggrin-2 was detected in the stratum granulosum with a granular labeling (arrows and insert) and in the stratum corneum. The polycarbonate filter–epidermal junction is indicated by a thin line. The mRNA and protein amounts corresponding to shc-RHE were arbitrarily set at 100. The error bars correspond to the S.D. calculated from independent experiments performed with keratinocytes from three different donors, one being duplicated for qRT-PCR (n=4) and each being duplicated for western blotting (n=6)