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The incorporation of [35S]sulfate and [3H] glucosamine into cetyl pyridinium chloride (CPC) precipitable glycosaminoglycans was determined in B16‐F10 cultured cells sorted with respect to DNA content. Incorporation into surface material was measured indirectly as the difference between the radio‐activity of control and trypsin treated cells. Approximately 80% of the total cellular [35S] sulfate labeled CPC precipitable material, but only 5% of that labeled by [3H]glucosamine, was removed by this mild trypsin treatment. Incorporation of [35S]sulfate into the trypsin removable surface material increased progressively from G1 to S to G2 + M in both long‐term (48 hours) and shortterm (1 hour) labeled cells, while the ratio of surface to total incorporated [35S]sulfate remained relatively constant. Incorporation of [35S]sulfate into total cellular glycosaminoglycans in long‐ and short‐term labeled cells increased as cells progressed from G1 to S to G2 + M; the incorporation of [3H]glucosamine into CPC precipitable material also increased progressively from G1 to S to G2 + M in long‐term labeled cells but was greater during S phase relative to G1 or G2 + M in short‐term labeled cells. The degree of sulfation of glycosaminoglycans as represented by the ratio of [35S]sulfate to [3H]glucosamine of double labeled cells was relatively constant in long‐term labeled cells but was increased during the G1 and G2 + M phases of short‐term labeled cells. Comparison of the degree of sulfation of short‐term with long‐term labeled cells suggests that highly sulfated glycosaminoglycans may be turned over more rapidly during G1 and G2 + M phases of the cell cycle.
Cytometry Part A – Wiley
Published: Nov 1, 1982
Keywords: ; ; ; ;
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