Figure 2.

Hexosaminidase Activity and Protein Expression in HEKHexABKO Cells Transfected with the HexB-HexA Construct

(A) MUG substrate was used to assess the hexosaminidase activity of cell lysates following DEAE column separation. ABKO cells showed no hexosaminidase activity above baseline, whereas WT HEK293 cells showed activity of all 3 isoforms of hexosaminidase. Following transfection with HexB-HexA, ABKO cells showed levels of hex activity equal to or greater than the WT HEK293 cells in all eluted fractions. (B) Protein expression analysis was conducted through western blot analysis, probing with anti-HEXA and anti-HEXB antibodies. Un-transfected HEKHexABKO cells showed no mature HexA or HexB expression, whereas ABKO cells transfected with the HexB-HexA construct (h.B2A plasmid) showed mature HexA and HexB expression levels.