Additional file 1 of CPT1A in AgRP neurons is required for sex-dependent regulation of feeding and thirst

Additional file 1: Figure S1. Validation of Cre-mediated recombination in AgRP neurons. a Scheme of the time-course of the experiment. b Bilateral injection of 400 nl of AAV9-EF1a-DIO-mCherry at a dose of 1.23 × 1013 gc/ml into the ARC of AgRP-Cre-ERT2 mice. c Representative histological slice of Cre-dependent mCherry expression in the ARC. Scale bar, 500 μm (top image) and 200 μm (bottom image). d Schematic of the Cre-mediated recombination product showing the floxed band (1030 bp) containing the LoxP sequences surrounding exon 4 of the Cpt1a gene. After Cre recombination, the product resulted in a 219-bp DNA fragment. e Scheme of the restriction sites of the restriction enzymes PstI and AatII that were used to separate Cpt1a amplicons from unrecombined genomic DNA. f Representative PCR analysis of genomic DNA from the liver, gonadal white adipose tissue (gWAT), brown adipose tissue (BAT), adrenal gland (AG), cortex, hippocampus (Hyp) and arcuate nucleus (ARC) treated with PstI and AatII enzymes in Cpt1aKO mice and control mice (g) FASTA analysis of the sequenced 219 bp DNA fragment extracted from the gel.