(A) Two abutted N2A cells expressing Cx36-EGFP GJ plaque. Junctional current was measured by applying voltage ramps from –10 to 10 mV to the cell-1 and measuring current response in the cell-2. Junctional conductance gj was obtained from the ratio –ΔIj/ΔVj. (B) Typical effect of hexanol and nonanol on Cx36-EGFP GJ conductance in N2A cells – gj is potentiated by hexanol and inhibited by nonanol. (C) Typical effect of hexanol and nonanol on Cx36C264S-EGFP GJ conductance in N2A cells – gj is inhibited by both hexanol and nonanol. (D) Summary of the effects of hexanol on GJ conductance of Cx36-EGFP and its mutants in HeLa and N2A cells. The effect of hexanol relatively to initial gj (indicated by dotted line) of Cx36-EGFP (indicated as C), Cx36C264S-EGFP, Cx36C92V-EGFP, and C36C264S/C92V-EGFP GJs was 3.4 ± 0.3 (n=9), 0.23 ± 0.06 (n=4), 0.88 ± 0.02 (n=4), and 0.36 ± 0.03 (n=8), respectively, in HeLa cells, and 3.3 ± 0.6 (n=6), 0.55 ± 0.06 (n=7), 0.96 ± 0.05 (n=6), and 0.38 ± 0.04 (n=8), respectively, in N2A cells. ***P<0.001, compared with Cx36-EGFP.