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Fig. 4. Comparison of wild-type and cys-less Cx43 hemichannel function in granulosa cells. (A) Propidium iodide (PI) uptake (red, central column) of Cx43-deficient granulosa cells infected with wild-type (WT) Cx43 (top row), cys-less Cx43 (middle row) or control vector (bottom row) under DCF conditions. Scale bar, 20 µm. (B) Quantification of PI uptake (percentage of EGFP-expressing cells loaded with PI) in regular ECS (baseline), in DCF solution or in DCF solution with 200 µM CBX in the three cell types, as shown under each group of bars. Data are expressed as mean ± s.e.m. of three to four different experiments under each condition. (C) Quantification of ATP release by Cx43-deficient granulosa cells infected with wild-type Cx43, cys-less Cx43 or control vector under regular ECS, DCF solution or DCF solution with 200 µM CBX. All data were normalized to baseline value (wild-type cells under regular ECS, broken line). n=4, 4 and 3 for the three cell groups, respectively. (B,C) ***P<0.001 versus control (two-way ANOVA and Bonferroni post-hoc test).

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