Fig. 5

Fig. 5 Inhibition of IGF signaling suppresses MFL development (A) Diagram illustrating the treatment regime for DMSO and the IGF1R inhibitor BMS-754807. (B–E) Confocal images of the lateral head at 6 dpf comparing the cartilage and lymphatic phenotypes of lyve1b:dsRed (green) sox10:EGFP (magenta) larvae treated at 4 dpf with either DMSO (B), 0.61 μM (C), 1.25 μM (D), or 2.5 μM BMS (E). Yellow arrowheads highlight the MFL. (F) Quantification of the length of the MFL comparing the DMSO control group (n = 16) with the groups treated with 0.61 μM (n = 16), 1.25 μM (n = 16), and 2.5 μM BMS (n = 21). (G and H) Diagram illustrating the workflow for the igfbp5b overexpression (G) and igfbp3 overexpression (H). (I–K) Confocal images of the lateral head at 6 dpf, comparing the lymphatic phenotypes between lyve1b:dsRed; col2a1a:Gal4-ERT; UAS:igfbp5b larvae treated at 3 dpf with either vehicle (I) or tamoxifen (J), and control lyve1b:dsRed; col2a1a:Gal4-ERT larvae treated at 3 dpf with tamoxifen (K). (L and M) Confocal images of the lateral head at 6 dpf, comparing the lymphatic phenotypes between lyve1b:dsRed embryos injected at the one-cell stage with either hsp70l:EGFP (L) or hsp70l:igfbp3 (M) following a heat-shock treatment at 4 dpf at 37°C for 2 h. Yellow arrowheads highlight the MFL. (N and O) RT-qPCR analysis of igfbp5b expression (N) or igfbp3 expression (O) at 4 dpf. (P) Quantification of the length of the MFL at 6 dpf, comparing the tamoxifen-treated igfbp5b expression group (n = 21) with the vehicle control group (n = 13) and the tamoxifen-treated control group (n = 11) or the hsp70:EGFP control group (n = 13) with the hsp70:igfbp3 group (n = 24). MFL, medial facial lymphatic vessel; OLV, otolithic lymphatic vessel. Scale bars: 50 μm. Data represent mean ± SD; ∗∗∗p < 0.001; ∗∗p < 0.01; ∗p < 0.05; ns, p > 0.05 from either one-way ANOVA with Dunnett’s multiple comparison tests for the assessment of significance (igfbp5b) or an unpaired t test with Welch’s correction (igfbp3).