Fig. 2. Wild-type, not Cx45.6 mutant, proteins form functional gap junction channels that mediate dye transfer. (A,B) Six days after retroviral infection with recombinant retroviruses RCAS(A)-Cx45.6, RCAS(A)-Cx45.6(D47A) and RCAS(A)-Cx45.6(P88S), the scrape-loading dye-transfer assay was performed using RD as a tracer dye (A,B) and Alexa488 (A) or LY (B) as transferring dyes. Bar, 40 µm. The extent of dye transfer was quantified by measuring the distance from scrape lines to the migrated front of cells stained with Alexa 488 (A, lower panel) or LY (B, lower panel). The data are presented as the mean±s.e.m.; n=5. (^*P<0.05; ^***P<0.001, in comparison with the non-Cx45.6-overexpressing control). Bar, 10 µm. (C) Six days after retroviral infection with recombinant retroviruses RCAS(A)-Cx45.6, RCAS(A)-Cx45.6(D47A) and RCAS(A)-Cx45.6(P88S), a parachuting dye-transfer assay was performed using Dil as a tracer dye and calcein as a transferring dye. The extent of dye transfer was quantified by measuring the area of calcein-fluorescence-stained cells (NIH image; lower panel). The data are presented as the mean±s.e.m.; n=3. ^***P<0.001, in comparison with the non-Cx45.6-overexpressing control. Bar, 10 µm.