Fig. 6.

Efficiency of aldolase knockdown by specific small interfering RNA (siRNA). A: Western blot analysis of aldolase expression levels was performed after 72-h transfection of HeLa cells with negative control siRNA (left lane), ALDO-A-selective siRNA-1 (middle lane), or ALDO-A-selective siRNA-2 (right lane). In each lane, 40 μg of total cell lysate proteins were loaded. Results of a representative experiment are shown. B: estimation of aldolase enzymatic activity in cell lysates after 72-h transfection of HeLa cells with negative control siRNA, ALDO-A-selective siRNA-1, or ALDO-A-selective siRNA-2. Data are presented as the time course of enzymatic cleavage of aldolase substrate fructose 1,6-bisphosphate (Fru 1,6-P₂), normalized to total protein concentration as described in materials and methods. Results of a representative experiment are shown. C: histograms show relative amounts of aldolase protein expression and enzymatic activity in siRNA-transfected cells. Normalized densitometric quantification of Western blot signals from A is shown (solid bars). Relative aldolase enzymatic activities from B were calculated as described in materials and methods and are shown (shaded bars).