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Fig. 9 PlcHR reduces endothelial cell numbers in zebrafish embryos.
(A?C) Acridine orange staining was used to indicate cell death at the location of endothelial cells in wildtype embryos injected with 2 ng of PlcHR at 6 hpi (arrows). Corresponding prime panels are enlarged areas showing cell death. (D?H) Zebrafish embryos expressing endothelial nuclear EGFP (Tg(Fli:nucEGFP) [46]) were utilized to assess PlcHR effects on endothelial cell numbers. As before, embryos at 48 hpf were injected with 2 ng of PlcHR, or PlcHR T178A, and compared with uninjected controls. To standardize the region used for counting nuclei, only those in the 8 ISVs immediately anterior to the cloaca were recorded [42]. (D,E,H) At 6, 12, and 24 hpi, there was no significant difference between PlcHR T178A-injected and uninjected animals. (C?F) Those injected with PlcHR showed a clear reduction in endothelial cell numbers by 6 hpi (P<.001). (H) Data taken from three independent experiments; n = 6, statistics performed by t-test.