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Figure S1 APETx2 inhibition of Nav 1.8 current is not use-dependent. Currents were recorded from acutely dissociated DRG neurones, with all bath solutions containing TTX. A series of 20 50 ms depolarizations from −70 to +20 mV were performed at the frequencies indicated (0.5–10 Hz). (A) A typical experiment at 0.5 Hz is shown. (B) Current inactivation was corrected by dividing the currents obtained with 1 μM APETx2 to that obtained without APETx2 at each pulse. This ratio is plotted as a function of the pulse number (1–20) for each stimulation frequency (n = 4 for each condition).

Figure S2 Sequence comparison of APETx2 with other sea anemone toxins. (A) Phylogenic tree, Nav, inhibitor of voltage-gated Na+ channels; K, inhibitor of K+ channels. Ae I from Actinia equina; Am II from Anthopsis maculata; ApA, ApB, from Anthopleura xanthogrammica; APETx1 and APETx2 from Anthopleura elegantissima; AsKS (kaliseptine), AsKC (kalicludines); BDS-I and BDS-II from Anemonia sulcata; BcIV from Bunodosoma caissarum; BgK from Bunodosoma granulifera; Da I from Dofleinia armata; PaTX from Entacmaea actinostoloides; Rp3 from Radianthus paumotensis; Sh1 and ShK from Stychodactyla helianthus. (B) Alignment of the novel sea anemone toxins to some of the classical Nav-targeting sea anemone toxins. Original references are cited in Honma and Shiomi (2006) and Shiomi (2009).

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BPH_1674_sm_FigureS1.pdf165KSupporting info item
BPH_1674_sm_FigureS2.pdf20KSupporting info item

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