Extended Data Fig. 2: Phenotypic Analysis of BCAT1 and BCAT2 Knockout Mice.

(a) Schematic of the genotyping PCR for Bcat1^−/− and Bcat2^−/− constitutive KO mice. The locations and sizes of the genotyping primers and PCR products are shown. (b) Representative genotyping PCR and Sanger sequencing results for Bcat1^−/− and Bcat2^−/− mice. (c) Validation of Bcat1^−/− KO by Western blot analysis of mouse brain and heart. HEK293T cells with BCAT1 and BCAT2 double knockout (DKO) and DKO cells with BCAT1 or BCAT2 overexpression (BCAT1^OE and BCAT2^OE) were analysed as controls. (d) Representative images for male and female mice of the indicated genotypes. (e) Representative H&E staining of epididymal adipose tissue in WT and Bcat2^−/− mice. Scale bar, 50 μm. (f) The organ weight relative to body weight is shown for heart, kidney and spleen of WT, Bcat1^−/− and Bcat2^−/− mice with the number (N) of samples shown. Results are mean ± SD and analysed by one-way ANOVA. *P < 0.05, n.s. not significant. (g) Hindlimb skeletal muscle and epididymal adipose weight of the indicated genotypes. Results are mean ± SD (N = 4 WT and 3 Bcat2^−/− mice for muscle, and 4 WT and 4 Bcat2^−/− mice for fat) and analysed by two-sided unpaired t-test. (h) Plasma AST and ALT levels in mice of the indicated genotypes. Results are mean ± SD (N = 5 WT and 4 Bcat2^−/− mice) and analysed by two-sided unpaired t-test. (i-k) Representative H&E staining of heart (i), kidney (j), and liver (k) from WT or Bcat2^−/− mice. Scale bar, 50 μm.

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