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Microtubules of secondary sprouts are selectively detyrosinated. (A-I′) Immunostainings using antibody detecting the glutamate amino acid of detyrosinated microtubules (dTyr) during primary (A-C) and secondary (D-I) sprouting in uninjected (A-F) and plcγ KD (G-I) Tg[fli1ep:EGFP-DCX] embryos, labelling all endothelial microtubules (DCX). C′,F′ and I′ are magnifications from boxed region in C,F,I, respectively. plcγ KD embryos (G-I′) show reduced primary sprouting, facilitating the visualization and quantification of dTyr signal specifically in secondary sprouts. Arrowheads indicate secondary sprouts. (J) Quantification of the dTyr/DCX signal intensities in primary sprouts of control MO-injected embryos, and secondary sprouts from plcγ MO-injected embryos. n=52 control primary sprouts from 18 embryos, n=24 plcγ KD secondary sprouts from 12 embryos, from one experiment. ****<0.00001 (Mann–Whitney test). Pictures are representative of 3 replicated experiments.
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