plxnd1 is required for the alignment of developing ISLVs to blood vessels. (A-H) Still images from time-lapse movies of ISLV growth between 60 and 70 hpf in lyve1b:DsRed; kdrl:EGFP wild type (A-D) and plxnd1nz75 (E-H) larvae showing blood vessels (green) lyve1b-expressing vessels (white) from Movies 1 and 2, respectively. Yellow arrowheads indicate the distal tip of each ISLV. (I,J) Traces of ISLV growth paths for wild type (I) and plxnd1 (J) (n=4). ISLVs were traced dorsally from when they left the horizontal myoseptum. (K) Quantitation of the percentage of time the ISLVs are aligned with a blood vessel (n=4). (L) Quantitation of the number of ISLV branching events (n=4). (M) Quantitation of the ISLV meandering index (n=4). *P<0.05 (Mann–Whitney test); data are mean±s.d. Scale bar: 50 µm. ISLV, intersegmental lymphatic vessel.
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