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. 2016 Apr 5;113(16):4326–4331. doi: 10.1073/pnas.1522372113

Fig. S4.

Fig. S4.

Fluorescence-based thermal-shift assays. (A) 15% (wt/vol) SDS/PAGE of purified recombinant wild-type and R30K mutated HA-His-RHD (amino acids 1–306) of RelA used for thermal-shift assays. (B, Left) Representative unfolding curves of HA-His-RHD WT and HA-His-RHD R30K over a temperature (T) range of 30–90 °C obtained under standard conditions (light gray and light blue) or in the presence of EMSA buffer components (dark gray and dark blue). (Right) Positive derivative [d(RFU)/dT] curves are shown. RFU, relative fluorescence unit. Midpoint temperatures of the protein-unfolding transition (Tm) are presented as bars. Values are mean ± SD of at least three independent measurements. (C) DNA-binding–dependent thermal stabilization of HA-His-RHD WT and HA-His-RHD R30K proteins in the thermal-shift assay. Results are presented as above.