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. 2004 Dec 23;102(1):93–98. doi: 10.1073/pnas.0408455102

Fig. 5.

Fig. 5.

Stimulation of the ASP expression and positive regulation of the dermal papilla-specific enhancer of the Agouti gene in vitro. Epidermal KCs and dermal fibroblasts were isolated from dorsal skin of C3H/HeJ mice at P2.5. KCs were cultured in the presence of 200 ng/ml BMP-4. RNA and proteins were harvested 24 h after treatment, and RT-PCR and Western blot analysis were performed. Dermal fibroblasts were cotransfected with plasmid vectors containing the 6.6-kb dermal papilla-specific enhancer that lies downstream the exons 1B and 1C of Agouti gene (pGL3-DPE-Lux) and one of the constitutively active BMP receptors. (A) Semiquantitative RT-PCR forβ-actin and ASP. Representative gels from one of three experiments each are shown. (B) Western blot analysis of 16-kDa ASP in the KC lysates after BMP-4 treatment. (C) Effects of cotransfection with vectors containing constitutively active BMPR-IA (Alk3QD) or BMPR-IB (Alk6QD) on transcription of the pGL3-Lux (negative control) or pGL3-DPE-Lux promoters (mean + SEM, **, P < 0.01, Student's t test).