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Fig. 4. Expression of p53, P-AKT, P-GSK3β and P-ERK1/2 in HK1.fos/ ${Delta}$ 5Pten phenotypes. Skin biopsies of keratoacanthomas (KA), papillomas (PAP), hyperplastic (HP) or hyperkeratotic (HK) epidermis, together with normal dorsal (N), anagen (aN) or ear skin (NE) were subject to western analysis. All HK1.fos/ ${Delta}$ 5Pten KAs expressed high p53 levels and phenotypic epidermis possessed low-level expression (mid panel) similar to normal controls (end panel). Conversely, p53 expression was undetectable in HK1.fos phenotypes (lanes PAP, HP and N) or hyperkeratotic K14.cre/ ${Delta}$ 5Pten^flx epidermis (end panel). HK1.fos/ ${Delta}$ 5Pten KAs expressed high but variable P-GSK3β levels, depending on tumour maturity ( ${Delta}$ 5Pten heterozygous KA 8898 versus homozygous KA 9593). However, KAs exhibited lower increases in P-AKT expression, which varied extensively with the degree of keratosis (Ka^*). Compared with total (t-) protein levels, HK1.fos epidermis exhibited low P-AKT and P-GSK3β expression (first panel), whereas P-GSK3β expression but not that of P-AKT increased in HK1.fos papillomas. Control K14.cre/ ${Delta}$ 5Pten^flx epidermis possessed elevated P-GSK3β and P-AKT expression (end panel). All hyperplastic phenotypes expressed elevated P-ERK1 and P-ERK2, including `normal' HK1.fos epidermis and HK1.fos papillomas in particular, which remained steady, if slightly reduced, in all KAs. β-Actin served as a loading control.

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