FIGURE 7.

Loss of UGDH intrinsic activity impacts cellular HA production. HEK293 cells were cotransfected with constructs encoding GFP (vector control) and human HAS3, or GFP and wild-type or mutant UGDH, or with HAS3 and each UGDH construct. After 48 h, conditioned media were analyzed for HA content, and cell lysates were analyzed by Western blot for UGDH, HAS3 (FLAG), or tubulin (control). HA was calculated in units of mass per volume of conditioned medium and normalized to protein content (in mg) of the whole cell lysates. The mean ± S.E. is plotted for quadruplicate measurements repeated three times. Statistical significance was evaluated by Student's t test; *, p < 0.01; **, p < 0.05, relative to HAS3 only. Western data are shown for the relevant lysates.