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Fig. 2. Hyaluronan production in Has2-transfected cells. (A) Nearly confluent
cultures were stained for cell-associated hyaluronan using the bHABC probe.
Wt, wildtype; M2, mock; A22, antisense; S29, sense cell line. Note the
elongation of some of the most intensely stained cells in the S29 sense line
(arrows). (B) Optical densities of the DAB signal, representing
cell-associated hyaluronan, were assayed in cultures of equal density to those
in (A) using microscopic image analysis. The bars show the s.e. of 10 randomly
taken microscopic fields. (C) Hyaluronan secreted into the growth medium by
the sense (S25-S30), antisense (A9-A25) and mock-transfected (M2) cells was
determined as fluorophore-tagged, enzymatically liberated disaccharides and
plotted as a function of cell density. Linear regression lines for the
antisense and sense cell lines are shown.