Figure 5

Schematic representation of the protocols used in the in vivo studies. (A) Zebrafish model of pseudoxanthoma elasticum (PXE). A total of 100 abcc6a^cmg52/cmg52 zebrafish larvae were exposed to 1 mM CSL525 at 3 days post fertilization (dpf) for 7 days, while another group of 100 larvae was kept under the same conditions without CSL525. After 7 days of treatment (10 dpf), the fish were euthanized, fixed in 4% paraformaldehyde, and stained with Alizarin Red S. (B) Mouse model of PXE. The abcc6^-/- mice were subcutaneously administered CSL525 at 15 mg/kg or 0.9% NaCl thrice weekly for eight weeks. A group of C57BL/6J wild-type mice, also receiving 0.9% NaCl thrice weekly, was included in the study. Each group consisted of five males and five females. All mice were maintained on a standard diet and sacrificed at 13 weeks of age, eight weeks after treatment initiation. (C) Rat model of calcinosis cutis. Calcinosis cutis was induced in 24 rats by sensitization with subcutaneous vitamin D (50,000 IU/kg on days 1, 2 and 3), followed by subcutaneous injections of FeCl₃ (0.15 mg) at each of the two ventral thoracic sites on day 4. CSL525 at 60 mg/kg (N = 12) or 0.9% NaCl (N = 12) was administered subcutaneously on a daily basis. A group of 6 healthy rats received subcutaneous 0.9% NaCl on days 1, 2, and 3, and 0.5 mL of 0.9% NaCl at each of the two ventral thoracic sites on day 4.