Fig. 1. Southern blot analysis of LINE-1 hypomethylation in bladder cancer cell lines. DNA isolated from leukocytes (left) was used as control. H: HpaII digestion; M: MspI digestion. Hypomethylation is detectable by the appearance of bands in the 1.4–3.5 kb range in the H lane. Results of densitometric quantification are depicted at the bottom of the figure.

Fig. 2. Cell cultures of urothelial carcinomas showing heterogenous and patchy nuclear staining with 5-mc antibody (original magnification × 400). (A) HT1376. A considerable portion of the cells is hypomethylated with granular staining pattern (in this picture staining score 2.3) (B) T24. Cells display mostly a stronger staining intensity compared to HT1376 (in this picture staining score 2.8).

Fig. 3. Urine samples of healthy controls demonstrating strong nuclear staining with 5-mc antibody (original magnification × 630). (A) Squamous cells with a homogenous fine granular staining pattern. (B) Basal transitional cell with dense nuclear staining. (C) Intermediate urothelial cells with a fine granular staining pattern comparable to that of squamous cells.

Fig. 4. Urine samples of two different patients (A + B) with urothelial carcinoma stained with 5-mc antibody (original magnification × 630). Cells of urothelial carcinoma (large arrows) display various degrees of destaining and often a patchy staining pattern. Compare to the strong homogenous staining of adjacent neutrophils (small arrows).

Fig. 5. Comparison of mean 5-mc immunostaining scores with 95%-confidence intervals in different groups, uninvolved epithelia and tumor cells. Hx: History.

Fig. 6. Comparison of methylation scores in tumor cells and matched uninvolved urothelial cells of individual patients. Each line displays the relationship between the methylation scores in the urothelial carcinoma and its paired normal epithelium.

Clinicopathological characteristics and 5-mc immunostaining scores of patients with urothelial carcinoma of the bladder

nd = not done (urine sample without cancer cells).

5-mc immunostaining scores in healthy controls, patients with benign disease of the bladder, patients with histologically confirmed urothelial carcinoma and patients with history of urothelial carcinoma but no evidence of recurrence