FIGURE 2.

Expression of His-tagged versions of AtFUT4.1 and AtFUT6 proteins in tobacco BY2 cells. A, dry weight of the cells was determined after 1, 3, 6, 9, 11, and 14 days of culture to monitor growth stages of the cells. The growth was steady until 9 days of culture and then stabilized, indicating a separation of cell division stage (3–9 days) from cell elongation stage (9–14 days). B, 7- and 14-day-old cultures of BY2 WT cells or transgenic BY2 cells were screened by Western blotting using anti-His₆ tag antibodies (Clontech). The cells were harvested and directly ground in liquid nitrogen before extraction with SDS-PAGE loading buffer, and then the proteins were separated on 10% gels. The protein bands detected had an estimated size of ∼60 kDa in agreement with the predicted size of AtFUT4.1 (∼61 kDa) and AtFUT6 (∼59 kDa) proteins. Marker sizes (kDa) are indicated on the left. MW, molecular mass; d, days.