Fig. 4. Effects of HAS2 and HYAL2 changes on HA synthesis and degradation.

a HeLa cells were transfected with low amount of mouse HAS2, high amount of BMR HAS2 (14-fold of mouse HAS2), or high amount (equal to BMR HAS2) mouse HAS2 plasmids. Two days after transfection, HA was purified from the media and analyzed by gel electrophoresis. b HeLa cells were transfected with equal amounts of shrew or SNM HAS2 plasmids and incubated for two days. HA was purified and run on an agarose gel. c Media containing 20 μg/mL commercial HMM-HA was incubated with confluent fibroblasts from each species. Two days after incubation, HA was purified from the media and run on an agarose gel. For each cell line, a sample without commercial HA was included to control the endogenous HA. d HeLa cells were transfected with either HYAL2 or HYAL1 of mouse, BMR, shrew, or SNM. Transfected cells were cultured in media containing 10 μg/mL commercial HA. Three days after incubation, HA was purified and run on an agarose gel. All experiments were performed three times with similar results.