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Figure 1


Fig. 1. Genomic structure of the human WNT16A and WNT16B isoforms. (A) WNT16A and WNT16B isoforms are produced by alternative expression of the first exon (1A or 1B) controlled by independent promoters (Fear et al., 2000). (B) First exon amino acid sequence alignment of the two WNT16 isoforms showing unique amino acid sequences encoded by each of the two first exons. (C-K) Protein and mRNA expression in normal human epidermis and basal cell carcinoma (BCC). (C) RT-PCR gel electrophoresis detection of WNT16B mRNA expression in normal human skin (NS), primary normal human keratinocyte culture (K), hairy human scalp skin (HS), primary dermal papillae culture (DP), BCC tumour samples (3 of 6 patients are shown here), blank (H2O), positive controls (plasmids WNT16A/B). WNT16B primers do not detect WNT16A template. M, molecular mass markers. GAPDH was used as a housekeeping gene control for all samples. (D) Gel densitometry quantification of WNT16B mRNA levels of NS, K, HS, DP and BCC samples. Each bar represents fold WNT16B activation (over NS as control) of mean ± s.e.m. with n number of samples indicated next to each bar. *P<0.05 and ***P<0.001 indicate statistically significant WNT16B mRNA activation over NS. (E) Immunoreactivity of WNT16 protein expression in normal human interfollicular epidermis showing cytoplasmic suprabasal staining pattern. (F) High power magnification of a portion within E showing WNT16 protein expression throughout the suprabasal layer (ss, stratum spinosum and sg, stratum granulosum) but discretely excluded the basal cell layer (sb, stratum basalae). (G-K) Immunoreactivity of WNT16 protein expression in BCCs and adjacent hair follicles. (H) High power magnification of a portion within G showing cytoplasmic WNT16 protein expression within BCC tumour mass (*) with increased WNT16 expression at the peripheral palisading advancing edge (arrowheads) of the tumour. WNT16 protein was also detected within the suprabasal layer in the epidermis above the BCC tumour. (I) Expression of WNT16 protein in a hair follicle adjacent to BCC tumours. Increased WNT16 protein expression was detected within the bulge region (b; J,K) and dermal papillae (dp; K) of the hair follicle.





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