Fig. S3.

Cd146 deficiency results in reduced claudin-5 expression and impaired pericyte recruitment. (A) Brain sections from cortex of mice at 4–6 wk were costained for CD31 (green) and claudin-5 (red) or ZO-1 (red) and analyzed by LSFM after being optically cleared by using organic solvents. Representative MIPs of 40 virtual single slices from Cd146^+/+ and Cd146^−/− mice are shown. (Scale bars: 50 μm.) (B) Quantification of the number of CD31⁺ capillaries expressing claudin-5 or ZO-1 and the MFI of claudin-5 or ZO-1 in CD31⁺ capillaries showed reduction of claudin-5 in capillaries of cortex from Cd146^−/− mice compared with those from Cd146^+/+ mice at P5 and 4–6 wk. (C) Brain sections from cortex of mice at 4–6 wk were costained for CD31 (green) and PDGFRβ (red) and analyzed by LSFM after being optically cleared by using organic solvents. Representative MIPs of 40 virtual single slices from Cd146^+/+ and Cd146^−/− mice are shown. (Scale bar: 50 μm.) (D) Quantification of the number of CD31⁺ capillaries of brain cortex from Cd146^+/+ and Cd146^−/− mice. No difference was detected. (E) Pericyte coverage was quantified by analyzing percent length of CD31⁺ capillaries as opposed to PDGFRβ⁺pericytes. The capillaries of brain cortex from Cd146^−/− mice showed a decrease in pericyte coverage (***P < 0.001). Data are from one experiment representative of three independent experiments with five mice per genotype, at least eight MIPs per mouse, and five random fields per MIP (B, D, and E).