Effects of S279 or S282 mutation on Cx43 expression and gap junction permeability in non-muscle cells. Representative western blots of lysates from HEK293 cells (A), which had been transduced with plasmids carrying rat nonspecific sequence (vector), wt-Cx43, S279A, S282A or S279A/282A genes for 48 hours, show the changes in the relative levels of pS279/282 and Cx43 expressions due to the mutations. Here, one blot membrane with 6 left bands and another membrane with 4 right bands from the same lysates were connected together to show all treatment groups. Some groups of the HEK293 cells were also treated with 5 μm 2-APB for 10 minutes, while others were stained with anti-Cx43 antibody to examine the subcellular distribution of Cx43 upon different treatments as indicated (B). Scale bar: 10 μm. The statistical data normalized by ?-actin depict the effects of different Cx43 mutants and 2-APB on S279/282 phosphorylation and Cx43 expression (C). N = 4 independent determinations for western blotting and immunostaining tests, respectively. In addition, the statistical data of LY uptake depict the effects of the different mutants on GJ permeability (D). N = 5–6 independent determinations for each bar. *P <0.05 and **P <0.01 vs. DMSO (0.1%); #P <0.05 and ##P <0.01 vs. vector, respectively.