DEV005041 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Phenotype of TAF4 inactivation in adult basal keratinocytes. (A-C) Hair cycle defects. Animals 4 weeks after Tam or oil injection as indicated. Note the hair loss at the snout and ventral regions. C shows a more extreme case of hair loss in a 18-month animal; compare with Fig. 2D. (D) Dry flaking skin on the feet of Tam-injected animals. (E,F) Hyperkeratinisation of the tail. E shows the tail from oil- or Tam-injected animals; F is a Haematoxylin-Eosin-stained paraffin section from the tail of a Tam-injected animal. (G,H) Histological analysis of Haematoxylin-Eosin-stained paraffin sections of tongue biopsies from oil- or Tam-injected animals 10 weeks after injection. (I) Transepidermal water loss (TEWL) from adult oil- or Tam-injected animals 10 weeks after injection. Two independent measurements were made from 14 oil- and Tam-injected animals. Mean values are shown with error bars where the value for oil-injected animals is set as 1 and the Tam-injected values are expressed relative this value.

• Supplemental Figure S2 -

  Fig. S2. Defective depilation-induced anagen. Histological analysis of Haematoxylin-Eosin-stained paraffin sections from skin biopsies taken 6-24 days after depilation. E, epidermis; D, dermis; HF, hair follicle; U, utriculi; DC, dermal cysts. The arrow indicates the basal membrane.

• Supplemental Figure S3 -

  Fig. S3. Increased phospho-CREB/ATF1 upon TAF4 inactivation. Sections of skin biopsies from oil- or Tam-injected animals were labelled with anti-phospho-CREB/ATF1 antibodies (#9191, Cell Signaling) or stained with Hoechst as indicated. BK, basal keratinocyte; E, epidermis; D, dermis; HF, hair follicle. Arrows indicate representative labelled cells.

• Supplemental Figure S4 -

  Fig. S4. Histological analysis of Haematoxylin-Eosin-stained paraffin sections from tumour sections. (A) Benign papilloma from an oil-injected animal. (B) Moderately differentiated squamous cell carcinoma from Tam-injected animal. Note irregular cell shape and internal keratinisation (arrow). (C) Large, locally invasive melanocytic tumour. (D) Squamous cell carcinoma infiltrated by melanocytes. Black arrow, keratin pearl; yellow arrow, pigmented melanocytes. (E) Lymph node infiltrated by melanocytes.

• Supplemental Figure S5 -

  Fig. S5. Immunodetection of TAF4 in keratinocytes and melanocytes. (A,B) To first confirm that the pigmented cells are melanocytes, sections from pigmented dermal tumours (A) and lymph nodes (B) were stained with the melanocyte-specific PNL2 antibody. Bright speckled cytoplasmic staining of the pigmented cells, but not the surrounding epidermal keratinocytes (A), or lymphocytes (B), is observed. Brightfield, PNL2 labelling, Hoechst staining and a merge of the PNL2 and Hoechst images are shown from left to right. (C) Tumour sections from oil- or Tam-injected animals as indicated. Left panel, immunostaining with anti-TAF4 antibody; centre panel, the Hoeschst-stained nucleus; right panel, the merged image. (D) Melanoma section from Tam-injected animal. Left panel is the brightfield image and the other panels are as described in C. TAF4 is absent from the keratinocyte-derived tumour from the Tam-injected animal, but is present in the melanocytes.

• Supplemental Figure S6 -

  Fig. S6. Changes in melanocyte gene expression. The expression of a series of genes involved in keratinocyte-melanocyte communication and of melanocyte-specific genes was investigated by quantitative RT-PCR on RNA from the skin of three oil- or Tam-injected animals. The value of the oil-injected is set as 1, and the Tam-injected values are expressed relative this value.

• Supplemental Table S1 -

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