(a) A pan-LCE1 antibody that recognizes LCE1 family members was generated using a synthetic peptide corresponding to the consensus His/Arg region, and an LCE1m-specific antibody was generated using a synthetic peptide that corresponds to a region in the carboxyl-terminus that is specific for this protein. (b) Immunofluorescent staining of back skin. Increased staining of both SPRR2 and the LCE1 antibodies was detected in the uppermost granular layers and cornified layers of LKO epidermis. Scale bars = 20 μm. (c) Protein from tape-stripped SC was subjected to immunoblotting. Both pan-LCE1 and LCE1m antibodies detected higher expression levels of LCE1 proteins in LKO SC. (d) Immunoelectron microscopic labeling of LCE1 family members in LKO stratum corneum and CEs isolated from WT and LKO mouse skin. Protein A-gold (10 nm) was used to detect bound antibodies on skin cryosections, while CE sections were probed with F(ab′)2 with ultrasmall gold, which was silver-enhanced for greater visibility. Scale bars = 100 nm. CE, cell envelope; His/Arg, histidine/arginine; LCE, late cornified envelope; LKO, loricrin knockout; SC, stratum corneum; WT, wild type.