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Fig. 8. Abnormal p53 activation in Cx43 ${alpha}$ 1KO PGCs. (A-C) Genital ridges from E11.5 wild-type and Cx43 ${alpha}$ 1KO mouse embryos were whole-mount stained using antibody against activated p53 (phospho-serine 15). PGCs (green) expressing activated p53 (red) are denoted by arrowheads. Quantitative analysis (C) revealed a marked increase in the expression of activated p53 in homozygous Cx43 ${alpha}$ 1KO PGCs. This analysis was carried out using nine Cx43 ${alpha}$ 1 +/+, 16 Cx43 ${alpha}$ 1 +/- and 11 Cx43 ${alpha}$ 1 -/- explants. (D) Assessment of PGC abundance in E11.5 genital ridges obtained from embryos of mice injected with ${alpha}$ -pifithrin or with vehicle alone (DMSO). With vehicle injection, homozygous Cx43 ${alpha}$ 1KO mouse embryos showed a significant reduction in PGC abundance. By contrast, following ${alpha}$ -pifithrin treatment there was no difference in PGC abundance in the wild-type versus heterozygous or homozygous KO embryos. This analysis included 22 Cx43 ${alpha}$ 1 +/+, 28 Cx43 ${alpha}$ 1 +/- and ten Cx43 ${alpha}$ 1 -/- ${alpha}$ -pifithrin-treated embryos, and 15 Cx43 ${alpha}$ 1 +/+, 18 Cx43 ${alpha}$ 1 +/- and ten Cx43 ${alpha}$ 1 -/- vehicle-treated embryos. (E) TUNEL labeling showed that following ${alpha}$ -pifithrin treatment the rate of apoptosis was similar in homozygous KO, heterozygous KO or wild-type PGCs. Note that the data are normalized to the frequency of TUNEL-positive PGCs detected in wild-type embryos. This analysis included five Cx43 ${alpha}$ 1 +/+, 37 Cx43 ${alpha}$ 1 +/- and 16 Cx43 ${alpha}$ 1 -/- ${alpha}$ -pifithrin-treated genital ridges. Scale bars: 25 µm.

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