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Fig. 3. HOPS expression in H-35 hepatoma cells. (A) Western blot analysis of HOPS in (Top) H-35 hepatoma cells arrested by double thymidine excess. Time 0: the cell at the end of treatment; 2-12 h: hours after addition of serum and release from cell cycle arrest. (Bottom) Western blot analysis of HOPS at different times after serum deprivation in H-35 hepatoma cells. 0-72 h: hours of serum deprivation; +1, +2 h: hours after addition of serum to cells. (B) An aliquot of H-35 cells was treated with propidium iodide and assayed by FACScan analysis to evaluate the cell cycle. (Top) H-35 cells not treated; (middle) time 0: H-35 cells following treatment with double thymidine; (bottom) 72 sd: H-35 cells starving for 72 hours following serum deprivation (sd). (C) Immunolocalization of HOPS in H-35 cells at different times (12-72 hours) after serum deprivation (12-72 hours) and 2 hours after addition of serum (+2 hours). Scale bars: 20 µm.

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