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Figure S1 (A) Amino acid sequence of synthesized peptides. (B) Analytical data obtained by MALDI-TOF spectrometry and by RP-HPLC. aPercentage of purity determined by HPLC using a buffer system [A = H2O (pH 2.5) and B = 100% CH3CN] with a gradient slope of 1% B per minute, at a flow rate of 1 mL·min−1 on a Vydac C18 column (5 μm particle size, 300 Å pore size). Detection at 214 nm. bObserved m/z-value compared with the calculated [M + H]+ monoisotopic mass. (C) Effect of hU-II or N-biotin-[Ahx0, Bpa3]-hU-II on Ca2+ mobilization using CHO cells co-expressing human UT receptors and a mitochondrial apo-aequorin protein. Data are expressed as a percentage of the response (bioluminescence) obtained with digitonin (50 μM) added to the same 96-well culture plate. Data represent the mean ± SEM of at least four independent assays performed in duplicate.

Figure S2 Enlargement of a nucleus from Figure 4D. Nucleus was stained with DRAQ5TM (blue colour). Left ventricular sections are double-stained for lamin A (red colour) and UT (green colour). This enlargement demonstrates the co-localization of UT (yellow colour, arrowhead) and lamin A.

Figure S3 Distribution of FITC-conjugated hU-II and URP in fixed untransfected CHO-K1 cells. Nuclei were stained with PI.

Figure S4 Distribution of FITC-conjugated hUII and URP in living (A) HeLa and (B) HEK-293 cells. Nuclei were stained with DRAQ5TM.

Figure S5 Internalization of 125I-hU-II and 125I-URP in CHO-K1 cells. CHO-K1 cells in 12-well plates were incubated for various periods of time in FBS-free media containing 0.2 nM 125I-hU-II or 125I-URP. Internalization was stopped by washing, and cells were then solubilized with 1 M NaOH, and the radioactivity was quantified using a γ-counter. Membrane-bound radioactivity was evaluated by incubation of CHO-K1 cells with 125I-labelled peptide (0.05 nM) for 30 s.

Appendix S1 Methods.

FilenameFormatSizeDescription
BPH_1710_sm_FigureS1.ppt172KSupporting info item
BPH_1710_sm_FigureS2.ppt235KSupporting info item
BPH_1710_sm_FigureS3.ppt6395KSupporting info item
BPH_1710_sm_FigureS4.ppt6468KSupporting info item
BPH_1710_sm_FigureS5.ppt249KSupporting info item
BPH_1710_sm_AppS1.doc56KSupporting info item

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