Supplementary Figure 1: Preparation of 40S–ABCE1 post-splitting complex.
From: Structure of the 40S–ABCE1 post-splitting complex in ribosome recycling and translation initiation
a, Outline of the facilitated splitting approach: 80S ribosomes were dissociated under high potassium (500 mM) and low magnesium (1 mM) conditions, followed by AMP‑PNP-dependent ABCE1 binding to the 40S subunit. Under re-association conditions (low potassium and high magnesium concentration (100 mM and 20 mM), free 40S subunits were allowed to rejoin with free 60S subunit. A representative profile of four individual preparations is shown. b, In ribosome profiles, the 40S–ABCE1 complex only appears in the 40S fraction if AMP‑PNP is present and ABCE1 is added under splitting-facilitating conditions. c, Pooled 40S ribosomal fractions show stoichiometric binding of ABCE1 to 40S subunits as analyzed by SDS-PAGE.
