DEV037168 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Growth of melanocytes from the harderian gland in vitro. (A) lacZ-stained harderian gland from a DCT-lacZ mouse at P0. DCT-lacZ-positive non-pigmented melanoblasts are shown. (B) Melanocytes cultured from a small piece of harderian gland taken from a P0 C56BL/6 mouse. Scale bars: 100 µm.

• Supplemental Figure S2 -

  Fig. S2. Growth of melanocytes from the harderian gland cultured on ST2 cells in vitro. (A) Every 12 hours, observations were made of a single, pigmented melanocyte derived from a P0 CAG-EGFP mouse. (Top rows) Bright-field image. (Bottom rows) Fluorescent image. (B,C) High-magnification views of an early (36 hours after plating, B) and late (264 hours after plating, C) culture showing that highly proliferative melanocyte lineage cells were not mature melanocytes. Scale bars: 50 µm.

• Supplemental Figure S3 -

  Fig. S3. Effect of various inhibitors on melanocytes of the harderian gland. (A-F) Number of colonies of melanocytes or DCT-lacZ-positive melanoblasts derived from P0 DCT-lacZ mice after 14 days of culturing 100,000 cells from the indicated cell source on ST2 cell monolayers in the presence or absence of the indicated inhibitors. (A) Epidermis, (B) dermis, (C) ear capsule, (D) vibrissa, (E) uvea, (E) harderian gland. A single set of experiments was performed simultaneously for all cell sources. The results of one set of experiments out of the two conducted are presented, and the same results were obtained in both sets. (G) Inhibitor-treated cultures. When melanocytes from harderian glands were cultured and exposed to the effective amounts of PD98059 or MET kinase inhibitor, their growth and/or differentiation was severely inhibited.