Extended Data Fig. 3: Generation of Alas1-/- immortalized preadipocytes using CRISPR-Cas9.
From: Haem biosynthesis regulates BCAA catabolism and thermogenesis in brown adipose tissue
a) mRNA levels of Alas1 in three representative Alas1 KO clones generated using three distinct gRNAs (n = 3 biological replicates per clone). b) Pparγ expression is not impacted by Alas1 deletion. c) Ucp1 levels are significantly reduced in all Alas1 KO clones (n = 3 biological replicates per clone for a-c). d) Protein levels of Alas1 and Ucp1 confirm loss of expression in screened clones. e) The relative abundance of numerous electron transport chain proteins is significantly reduced in Alas1 KO brown adipocytes compared to WT (n = 4 biological replicates per genotype). Data shown as mean ± SD. p values vs. WT; one-way ANOVA with multiple comparisons and a Tukey’s post-test.
