JCS007104 Supplementary Material

Files in this Data Supplement:

• Supplemental Table S1 - Adobe PDF

• Supplemental Figure S1 -

  Fig. S1. Conditional knockout of Miz1 POZ/BTB domain in keratinocytes. (A) The N-terminal POZ/BTB domain of Miz1 is encoded by exons 3 and 4 (E3, E4), which were flanked by loxP sites (black triangles) by homologous recombination. Arrows indicate primers used for PCR or RT-PCR, respectively. (B-D) Immunohistochemical staining of mouse skin with hair follicles in anagen (B) or in telogen (C) using the antibody 10E2 against Miz1. Independent of the hair cycle stage Miz1 was present in the nuclei of basal and some suprabasal cells of the hair follicle epithelium. (D) Control slide stained without first antibody.

• Supplemental Figure S2 -

  Fig. S2. Histology of the back skin from K14Cre^-/Miz1^lox/lox and K14Cre⁺/Miz1^lox/lox mice from different embryonic and postnatal stages as indicated. The irregular hair follicle arrangement and epithelial thickening at the inter- and intrafollicular epidermal transition is first detected between day P4 and P7. The magnification is equal in all pictures. Bar, 200 μm.

• Supplemental Figure S3 -

  Fig. S3. Analysis of gene expression and cell proliferation in isolated keratinocytes. Keratinocytes from control or from K14Cre⁺/Miz1^lox/lox mice were isolated and incubated with or without 100 pM TGFβ1. (A) Miz1 mRNA was almost undetectable when primers 3 and 4 were used (see Fig. 1B). Myc was downregulated by TGFβ1 irrespective of the genotype, and the mRNA of Myc-activated genes, such as nucleolin was also decreased. The induction of p15ink4b by TGFβ1 was slightly reduced in Miz1^ΔPOZ/ΔPOZ keratinocytes compared with control keratinocytes. (B) Expression of the indicated genes revealed a reduced response to TGFβ1 in Miz1^ΔPOZ/ΔPOZ keratinocytes compared to control keratinocytes, similarly to p15ink4b. This was also confirmed for some genes by RQ-PCR from an independent preparation of keratinocytes (D). (C) Examples of genes, which show no differential response to TGFβ1 when Miz1^ΔPOZ/ΔPOZ keratinocytes were compared with control keratinocytes.