Biological solution conditions and flanking sequence modulate LLPS of RNA G-quadruplex structures

Thermal denaturation of 3′-flanking sequences in the absence and presence of polyamines. Sequences were renatured in the (A) absence or (B) presence of 0.4 mM spermidine and 0.3 mM spermine. T_m is the temperature with a minimum in the first derivative. Microscopy fields are shown with corresponding sequences. All samples are in the background of 10 mM HEPES (pH 7.5) and K₁₅₀N₁₀M_0.5. Melts were monitored at 294 nm with a heating rate of 0.5°C/min. (G₃A₂)₄ is shown in orange as a reference point for the early melting transition in both polyamines.