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Fig. 3. PN-1-mediated inhibition of thrombin activity affects proliferation of cultured hair follicle dermal cells. (A) Growth of wild-type and PN-1^–/– DP cell in medium containing 10% FCS. Cells were plated at 200 cells/mm² and average cell density from three dishes was calculated at different time points after plating. (B) Effect of hirudin-mediated inhibition of thrombin in serum-supplemented culture medium on cell proliferation. Hirudin (100 µM) was added 24 hours before a pulse of BrdU (10 µM, 2 hours). ^***Significant difference from wild-type cells (P<0.001). (C) Effect of PN-1 on volume increase. Cells transferred to medium containing 1% FCS were blocked in S phase by aphidicolin (1 µg/ml) for 24 hours and then cultured for an additional 24 hours with or without recombinant PN-1 (rPN-1; 15 nM) in the absence or presence of RAP (1 µg/ml). Change in mean volume was calculated for non-dividing cells in three dishes for each treatment. ^**Significant difference between PN-1^–/– and wild-type cells (P<0.01); ^*significant difference between PN-1^–/– cells with or without rPN-1 (P<0.05).

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