FIG 10.

Reduced acylceramide levels in ALDH3A2 KO human immortalized keratinocytes. (A) ALDH3A2 KO keratinocytes were generated using the CRISPR/Cas9 system. The exon structure (red, coding sequence; blue, untranslated regions) of human ALDH3A2 and the nucleotide sequences of WT and ALDH3A2 KO cells (KO clones 1 and 2) around the guide RNA target sequences (light blue) and the PAM sequences (magenta) in exon 4 are shown. The green nucleotide indicates a missense mutation. (B and C) Lipids were extracted from two control cells and two ALDH3A2 KO cells on day 14 of differentiation, and acylceramides (B) and ceramides (C) were analyzed via LC-MS/MS. The quantities of each lipid species (according to chain length and saturation) and of total lipids are shown in the left and right panels, respectively. Values presented are means ± SD (*, P < 0.05; **, P < 0.01; Tukey’s test) from three independent experiments.