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Fig. 5. Expression of Cx43 mRNA (A) and protein (B) in J774 cells. (A) Relative expression of Cx43 mRNA in ATP-s and ATP-i cells, obtained by amplified Cx43 and GAPDH RT-PCR products. The graph represents the mean±s.d. values for the ratio between Cx43 and GAPDH bands (n=5). The insert shows the agarose gel of a semiquantitative RT-PCR experiment. Mock is used as a negative control. There were no significant differences between ATP-s and ATP-i cells. (B) Western blot showing that ATP-s and ATP-i cells express Cx43 protein (43 kDa arrow). Liver and brain homogenates were used as negative and positive controls, respectively. Each lane was loaded with 100 µg of protein and separated by 10% SDS-PAGE before transfer to nitrocellulose membrane. The histogram represents the mean±s.d. (n=5) of relative densitometry values for Cx43 labeling taking the rat-brain homogenate as reference. The intensity of the bands in ATP-s and ATP-i cells are not significantly different from each other. The asterisk indicates that murine brain homogenate expressed significantly more Cx43 than the other cells (P<0.05).
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