Table 2 Biophysical characterisation data for the sequences of the G-rich ILPR variants

Sequence Flank₁-G-tract-Loop₁-G-tract-Loop₂-G-tract-Loop₃-G-tract-Flank₂		UV spectroscopy melting, annealing temperatures (°C)				CD	TDS	QGRS score
		295 nm		260 nm		CD	TDS
		T_m	T_a	T_m	T_a	Structure
1G	ACAGGGGTGTGGGGACAGGGGTGTGGGGACA	76 ± 0.6	60 ± 0.0	77 ± 0.6	60 ± 0.6	G4	G4	63
2G	ACAGGGGTGTGGGCACAGGGGTGTGGGCACA	56 ± 0.0	50 ± 0.0	56 ± 0.6	51 ± 0.6	Mix	Mix	42
3G	ACAGGGGTGTGAGGACAGGGGTGTGAGGACA	30 ± 0.0	27 ± 0.6	31 ± 0.6	28 ± 0.6	G4	Hp	21
4G	ATAGGGGTGTGGGGATAGGGGTGTGGGGATA	79 ± 0.6	60 ± 0.6	79 ± 0.6	60 ± 0.6	G4	G4	63
5G	ATAGGGGTGTGTGGATAGGGGTGTGTGGATA	nt	nt	52 ± 0.0	37 ± 0.6	Hp	Hp	21
6G	ACAGGGGTCTGGGGACAGGGGTCTGGGGACA	82 ± 0.7	56 ± 0.6	81 ± 0.6	57 ± 0.7	G4	G4	63
7G	ACAGGGGTCTGAGGACAGGGGTCTGAGGACA	nt	nt	51 ± 0.0	48 ± 0.6	Hp	Hp	21
8G	ACAGGGGTCCGGGGACAGGGGTCCGGGGACA	80 ± 0.7	60 ± 0.6	81 ± 0.6	60 ± 0.0	Mix	Mix	63
9G	ACAGGGGTCCTGGGGACAGGGGTCCTGGGGACA	nt	nt	57 ± 0.6	51 ± 0.6	Hp	Hp	62
10G	ACAGGGTCCTGGGGACAGGGTCCTGGGGACA	nt	nt	54 ± 0.6	51 ± 0.0	RC	Hp	42
11G	ACAGGGGTCCCGGGGACAGGGGTCCCGGGGACA	nt	nt	68 ± 0.0	63 ± 0.0	Hp	Hp	62

ILPRG sequences and their labels (1G-11G). Putative loops are underlined. All biophysical analyses were performed at pH 7.0. Most prominent UV melting, annealing and hysteresis temperatures, and structure characterisation from the CD and thermal difference spectra (TDS), and QGRS Score³⁰. UV and CD spectroscopy were repeated in triplicate and data are shown as mean ± SD (n = 3). G4 = G-quadruplex; Mix Mixed species, Hp hairpin, nt no transition observed.

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