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Figure 2. Microinjection of dvl2-D2 mRNA increases the number of phospho-H3 positive cells in the pronephros. Embryos were injected with dvl2-D2 plus membrane bound green fluorescent protein (GFP) mRNAs at the eight-cell stage, grown to stage 21/22, then fixed. Fluorescent in situ hybridization (FISH) was performed in the green channel to detect the pronephros (panels A and B), tyramide enhanced immunochemistry in the red channel to detect phospho-H3 (panels C and D), and tyramide enhanced immunochemistry to detect GFP in the far red channel (shown in blue). A,C,E,G: The left column shows different channels (A,C,E) of a control sample with a merge of these presented in panel G. The right column shows a similar data set for a dvl2-D2 plus GFP mRNA-injected pronephros. F,H: Due to the sequence of development some of the phospho-H3 signal also shows up in panels E and F, but is double stained and appears as magenta in panels G and H allowing the clear distinction of phospho-H3 and GFP. |
