Figure 8.
Hexokinase interacts with one or more 14-3-3 proteins in human neuroblastoma cells. (a) Acrylamide gels were loaded with 50 μg total protein per lane extracted from SH-SY5Y-APPSw cells after a 48 h exposure to FDA-approved drugs lumacaftor or conivaptan. Western blots were probed with antibodies to the 14-3-3 conserved core, hexokinase, or GAPDH. (b) Mean ± SD band intensities of hexokinase lanes (N = 3) normalized to the corresponding 14-3-3 band. (c) Antibody to the 14-3-3 conserved core was used for immunopulldown of 14-3-3 complexes from neuroblastoma cells either treated for 48 hr with drugs or untreated. Gel lanes were loaded with protein recovered from coated magnetic beads and electrophoresed, and western blots were probed to detect hexokinase or 14-3-3 proteins. (d) Histogram of hexokinase recovery (signals on western blots as in (c)), normalized to 14-3-3; the band signal is plotted for mean ± SD band intensity ratios of replicates. Significances according to two-tailed heteroscedastic t-tests (b,d), each N = 4: * p = 0.05; ** p < 10−2; *** p < 10−3.