Western blots showed that the total Cx43 content of rat heart slices decreased with the increasing number of days in culture (A). Only freshly cut slices (0 day) contained all three phosphorylated forms of Cx43. Quantification (B) showed an approximately 70% decrease in total Cx43 expression after 1 day, decreasing 13% in rat slices after further days in culture. FP/PP, fully/partly phosphorylated forms of Cx43; 0P, dephosphorylated form of Cx43. n=4 at each time point. Rat embryonic heart slices remained viable for longer in culture. Cx43 labeling of embryonic heart slices after 6 days in culture (C) showed positive gap junctional labeling similar in appearance to that observed in whole embryonic rat heart sections (D). One representative picture from 4 slices (from 4 different hearts) at each time point is presented. Bar markers = 100 μm. Western blot analysis (E) demonstrated that even after 35 days in culture levels of Cx43 remained similar to those observed in freshly-cut (day 0) slices and that all three phosphorylated forms were present.

Figure 4: Different Cx43 expression between adult and embryonic heart slices

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