Fig. 4: Perturbations of VEGF-C to VEGFR3 signaling influence the side-branching and space-filling efficiency of lymphatic capillary networks. | Nature Communications

Fig. 4: Perturbations of VEGF-C to VEGFR3 signaling influence the side-branching and space-filling efficiency of lymphatic capillary networks.

From: Self-organized and directed branching results in optimal coverage in developing dermal lymphatic networks

Fig. 4

A–C LYVE1 stained ventral ear pinna dermis of control (n = 12 ear pinna, representing 6 mice) or soluble VEGFR3 receptor (VEGF-C ligand trap) (n = 14 ear pinna, representing 7 mice) treated (from P11 to P21) mice. The dashed line indicates the ear pinna boundary. The boxed regions are shown as magnified images. B Mean +/− SD of the segment number (p < 0.0001), normalized to the average of controls (set as 1), and median segment length (p < 0.0001) of control (n = 12 ear pinna) and sVEGFR3 (n = 14 ear pinna) treated mouse ear pinna ventral dermis lymphatic capillaries. C Quantification of the spatial fluctuations at P21 in control (n = 12 ear pinna) or sVEGFR3 (n = 14 ear pinna) treated mice, showing enhanced fluctuations in the latter (slope \(\alpha=0.56\)). D, E consistent with simulations with abolished branching from P11 (as well as random pruning, see Supplementary Information Theory Note for details). F–H LYVE1-stained ventral ear dermis of control (n = 8 ear pinna, representing 4 mice) and Vegfc+/− mice (n = 8 ear pinna, representing 5 mice). The yellow dashed line indicates the ear pinna boundary. The boxed regions are shown as magnified images. G Mean +/− SD of segment number (p < 0.0001), normalized to the average of controls (set as 1) as in (B), and median segment length (p = 0.0003) of lymphatic capillaries in control (n = 8 ear pinna) and Vegfc+/− (n = 8 ear pinna) mice. H Quantification of the spatial fluctuations at P21 in wild-type (n = 8 ear pinna) and Vegfc+/− (n = 8 ear pinna) mice, showing enhanced fluctuations in the latter (slope \(\alpha=0.65\)), I, J consistent with simulations with abolished side-branching and decreased tip-branching (to 25% of its WT value, based on (G). K LYVE1-stained ventral ear dermis of control (n = 6 ear pinna, representing 4 mice) and Clp24ΔEC mice (n = 4 ear pinna, representing 2 mice). Clp24ΔEC was deleted at P8. See Supplemental Fig. 9A for further examples. L Mean +/− SD of the segment number, normalized to the average of controls (set as 1), and median segment length of control and Clp24ΔEC mouse ear pinna ventral dermis lymphatic capillaries upon 4-OHT mediated Clp24 deletion at p8 (control n = 6 ear pinna, representing 4 mice and Clp24ΔEC n = 4 ear pinna, representing 2 mice), P11 (control n = 5 and Clp24ΔEC n = 6 ear pinna, representing 3 mice each) or P13 (control n = 3 and Clp24ΔEC n = 4 ear pinna, representing 3 mice each). For comparison of segment number in controls and Clp24ΔEC, p = 0.002 (Clp24 deletion at P8), p < 0.0001 (at P11), and p = 0.003 at (P13), whereas for comparison of segment length p = 0.003, p < 0.0001, and p = 0.001, respectively. M Close-up of simulations for WT and Clp24ΔEC mouse (150% branching rate compared to WT), showing good qualitative agreement with the data. Scale bars for overview and magnified images in (A) and (F) are 1 mm and 200 μm, respectively, and for (K) 200 μm. In (B), (G), and (L), two-sided Welch’s t test was used for measuring statistical significance. Source data for Fig. 4B, C, G, H, L are provided as a Source data file.

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