Diarylheptanoids from Alpinia officinarum Cause Distinct but Overlapping Effects on the Translatome of B Lymphoblastoid Cells
Figure 2
Polysome sedimentation profiles of fractions isolated from control BJAB cells and BJAB cells treated with AO-0001, AO-0002, or AO-0003. The cytosol fraction was layered onto a 0.5 M–1.5 M linear sucrose gradient (prepared by Gradient Master 107, Biocomp) in LSB (20 mM Tris-HCl pH 7.0, 10 mM NaCl, 3 mM MgCl2); the gradient tube was centrifuged at 47,000 rpm in a Hitachi SW55Ti rotor for 70 minutes. A Piston Gradient Fractionator (Biocomp) equipped with an absorbance monitor (254 nm) was used to separate each gradient into nine fractions (0.5 mL/fraction). The absorbance of each 9th fraction (0.5 mL) could not be measured because, in each case, the 9th fraction was retained at the bottom of the ultracentrifuge tube. Positions of ribosomal 40S and 60S subunits and 80S monosomes are indicated by arrows.
