4.8 Electrophoretic mobility shifts assay (EMSA)
EMSA were performed as previously described (Lei et al., 2020).
Constructs for MBP-BES1 (Li et al., 2020) were transformed intoEscherichia coli strain BL21 (DE3) and recombinant proteins were
induced by IPTG. MBP-BES1 was purified using Amylose resin (Cat no.
E8021; New England Biolabs, Ipswich, MA, USA). Purified MBP-BES1 was
incubated with the probe listed in supplemental table, the mixture was
reaction in binding buffer (25 mM HEPES-KOH, pH 8.0, 50 mM KCl, 1 mM
dithiothreitol and 10% glycerol) for 30 min and shift in polyacrylamide
gel. The shifted probe and protein were tested with EMSA Kit (Thermo
Fisher).