Fig. 6

Cx26 mutants failing to complex with NANOG or FAK/NANOG disrupt the self-renewal capacity of TNBC CSCs. a, b After transfection of plasmids expressing GFP-fused Cx26 wild-type and mutant proteins, immunoprecipitation was performed using a GFP antibody in MDA-MB-231 and PDX TN-1 cells, and the resulting immunoprecipitates were probed with FAK, NANOG, and GFP antibodies. c Stem cell frequencies determined by limiting dilution sphere-forming assays indicate that the expression of the D66H-RFP mutant Cx26 in MDA-MB-231 CSCs significantly reduced the stem cell frequency compared with the expression of the wild-type Cx26-RFP and empty vector control. d D66H-GFP or G59A-GFP mutant Cx26 expression in MDA-MB-231 parental cells failed to increase stem cell frequency compared with the expression of the wild-type Cx26-GFP (*p < 0.05, **p < 0.01, ***p < 0.001). All the error bars indicate the range between the upper and median levels