JCS047274 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. AhR expression in normal skin and during wound healing. Wounds were made in wild-type mice and tissue dissected at 3 and 5 days. Tissues were fixed and analyzed by immunofluorescence using a rabbit anti-AhR primary antibody and an Alexa Fluor 488-labeled secondary antibody. Total cell extracts were also obtained from basal skin and day 3 wounds and analyzed by western blotting using an AhR primary antibody. The expression of β-actin was detected as loading control. AhR expression was normalized by β-actin levels. No significant differences were observed in AhR expression during wound healing and with respect to normal skin.

• Supplemental Figure S2 -

  Fig. S2. Identification of keratinocytes in skin explants by immunoflourescence. Skin explants were grown for 48 hours and stained with a keratinocyte-specific antibody against keratin 14 labeled with TRITC (AF 64 from Covance). Nuclei were stained with DAPI. Note that cells emigrating from the explants were positive for keratin 14 and therefore of epithelial phenotype.

• Supplemental Figure S3 -

  Fig. S3. Quantitative real-time RT-PCR analyses of Tgfbr1 and Tgfbr2 mRNA expression in AhR^+/+ and AhR^−/− primary keratinocytes. Total RNA was purified from keratinocytes of either genotype and gene expression analyzed by quantitative RT-PCR following published protocols (Gomez-Duran et al., 2008a). Tgfbr1 and Tgfbr2 expression were normalized by that of β-actin. The oligonucleotide primers used are indicated at the bottom of the Figure. Tgfbr1 primers: Forward, 5′-GGC GAA GGC ATT ACA GTG TT-3′; Reverse, 5′-TGC ACA TAC AAA TGG CCT GT-3′; Tgfbr2 primers: Forward, 5′-GCT TGG CCA GAA AGA CAG AC-3′; Reverse, 5′-CAC TCC ACA AGC TGT CTC CA-3′.