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Fig. 8 The effect of PORCN inhibitors on chondrogenesis in zebrafish embryos. (A) Schema of experimental design showing that PORCN inhibitors were added into zebrafish cultivation media at 0, 12, or 24 hpf to identify the effect of inhibitors during early embryogenesis or later during chondrogenesis in vivo. All embryos were collected at 5 dpf. (B) External morphology of zebrafish at stage 5 dpf. Increased concentration of LGK974 induced progressive shortening of the longitudinal axis of embryos in animals treated at 24 hpf. Scale bar = 2 mm. (C) Jaw cartilages of 4 dpf transgenic Sox10:DsRed zebrafish were shorter and thicker after cultivation with C59 or LGK974 (1 μM concentration added at 12 hpf) with a disrupted pattern of chondrocytes alignment. Scale bars: Lower view = 100 μm, details = 20 μm. (D) Analysis of palatoquadrate length, palatoquadrate width, and Meckel’s cartilage following C59 and LGK974 treatment. Statistical analyses were performed using the Mann–Whitney test (n = 6). Abbrevation: PORCN, Porcupine.