Fig. 6. Dlx3 is an indispensable regulator for the expression of follicular-specific structural and transcription factors. (A) Expression of Hoxc13, Gata3, and A13 and A15 hair keratins were significantly reduced in the K14Cre;Dlx3^Kin/f follicles at P1 and P9 anagen stages. White arrowheads indicate expression areas of each factor. Scale bar: 50 µm. (B) Luciferase reporter assays of the 1.2 kb K35 promoter and mutants 2, 6 and 7 sites in the absence and presence of Dlx3 co-expression. (C) (Left) ChIP assays were performed in primary and transfected mouse hf cells using anti-Dlx3 and anti-V5 antibodies, respectively. Triangles indicate the putative binding sites in the promoters for hair keratins (K32 and K35) and Hoxc13, which are conserved between human (white) and mouse (black). Each number in the triangles correspond to the sequences analyzed by EMSA (see Fig. S3B in the supplementary material). The PCR amplified areas are indicated by black lines in the mouse promoters for hair keratins (K32 and K35) and Hoxc13. (Middle) Gel images for each ChIP assays; the specificity of the amplified fragments was verified by sequencing. (Right) Specificity of the ChIP assays determined by both control IgG antibody and a set of PCR primers for Gapdh.