Additional Supporting Information may be found in the online version of this article.

IJC_25629_sm_suppfig1.tif3018KSupporting Figure 1 Normal skin from a total vulvectomy analysed in our study, without any sign of lesion (Supplemental fig. 1a); heterogeneous pattern of TSP-1 expression, with positive and negative areas coexisting with hypermethylation of the gene (case 25) (Supplemental fig. 1b).
IJC_25629_sm_suppfig2.tif1360KSupporting Figure 2 Reverse line blot (RLB) of the LS not associated with vulvar SCC; the 37 types tested in RLB are listed in the Methods. High-risk type HPV16, HPV18, HPV58 and a combined infection of low-risk type HPV6 and high-risk type HPV59 were found.
IJC_25629_sm_suppfig3.tif3063KSupporting Figure 3 MS-PCR of TSP-1 gene in LS not associated with vulvar SCC. Both the unmethylated (U) and methylated (M) PCR products are shown for each case. The signal of the methylated band is less intense than the controls. DNA from normal lymphocytes (NL) and in vitro-methylated DNA (IVD) are used as a positive PCR control for the unmethylated and the methylated reactions, respectively; H2O: Negative control.
IJC_25629_sm_suppfig4.tif7114KSupporting Figure 4 Bisulphite-sequencing analysis of RASSF1A, RASSF2A, p16, TSP-1 and MGMT promoter methylation status in normal vulvar skin, isolated lichen sclerosus, and paired associated lichen sclerosus and vulvar tumor. For each clone the methylation status of analysed CpG sites is shown (open and filled circles represent unmethylated and methylated circles, respectively). Densely methylated clones are present in associated lichen sclerosus and vulvar tumours. Black arrows represent the location of the methylation specific PCR (MSP) primers. The location of each CpG site relative to the transcription initiation site is shown by a vertical bar.
IJC_25629_sm_supptable1.doc43KSupporting Table 1.
IJC_25629_sm_supptable2.doc31KSupporting Table 2.
IJC_25629_sm_supptable3.doc39KSupporting Table 3.

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