This PDF file includes:

• Fig. S1. Cryo-EM of the undocked WT INX-6 hemichannel in a nanodisc.
• Fig. S2. Comparison of INX-6 channels between docked and undocked forms focusing on extracellular domains and the pore pathway.
• Fig. S3. Sequence alignment of INX-6 and docking interface residues.
• Fig. S4. Cryo-EM of the undocked WT INX-6 hemichannel in detergent prepared by GraDeR.
• Fig. S5. Cryo-EM of INX-6ΔN in an undocked hemichannel form in a nanodisc.
• Fig. S6. MD simulations of the undocked INX-6 hemichannels.
• Fig. S7. Fluorescent micrographs of INX-6 distribution on Xenopus oocytes.
• Table S1. Summary of data and statistics of the undocked INX-6 hemichannel structure determination.
• Reference (69)

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