Figure 1. CD36 distribution in human and mouse hearts and creation of EC- and CM-Cd36^–/– mice.

(A) CD36 antibody staining of CMs and blood vessel ECs from human and mouse hearts. Original magnification, ×20. (B) Male mouse heart suborgan fractionation showing Cd36 mRNA levels in different cell types (n = 12–18). Data represent mean ratios normalized to CM-Cd36^–/– (set at 1.0). (C) Genomic structure of murine Cd36, the targeting vector, and the mutated allele. Gray boxes represent the numbered murine Cd36 exons. Red arrows indicate transcription orientation. Black bent arrows indicate the translation start site. PCR primers (blue arrows) were chosen to differentiate between the WT genomic allele and the homologously recombined allele. (D) PCR of tail genomic DNA (using primer 1) and (E) heart tissue DNA (using primer 2) from Cd36^fl/fl, EC-Cd36^–/–, and CM-Cd36^–/– mice. (F) PCR product sequencing showing ablation of exons 3 and 4 of Cd36 gene after Cre-mediated recombination. (G) Heart Cd36 mRNA in EC-Cd36^–/– and CM-Cd36^–/– mice quantified by qRT-PCR using primer 3 (n = 5–6, mean ± SD). Data are corrected for 18S rRNA and normalized to Cd36^fl/fl controls. Immunoblots show CD36 in EC-Cd36^–/– and CM-Cd36^–/– mouse hearts. (H) Immunoblots of CD36 in EC-Cd36^–/– muscle, WAT, BAT, and liver. *P < 0.05, ^#P < 0.01 and ^§P < 0.001 compared with Cd36^fl/fl controls; 1-way ANOVA with Dunnett’s multiple comparisons test.