Extended Data Fig. 8: ALBA5 binds ssRNA in vitro and in vivo.
From: ALBA proteins confer thermotolerance through stabilizing HSF messenger RNAs in cytoplasmic granules
a, Coomassie blue staining of His-tagged ALBA5 purified from E.coli. Data are representative of five independent experiments with similar results. b, EMSA results showing ALBA5 binding to ssRNA probes and competition by unlabeled ssRNA probes. 3′-Fam-labeled substrates (80 nM) were incubated with increasing concentrations of ALBA5. The protein concentrations in lanes 1, 2, 3 and 4 were 0, 1, 2 and 3 μM, respectively. For competition analysis, 32 μM of unlabeled substrate was used as a cold probe and incubated with 3 μM of ALBA5-His and 80 nM of 3′-Fam-labeled substrates. For EMSA, at least three independent biological replicates were performed, and representative results are shown. c, Principal component analysis (PCA) plots of ALBA5 binding clusters in 5-day-old seedlings under control (22 °C) and heat stress (39 °C for 50 min) conditions.
