From 5 to 25 weeks of age, females were weighed weekly without monitoring the estrous cycle as KO and KIKO do not exhibit a normal estrous cycle34 (link),35 (link). At the end of 25 weeks, body composition was measured using an EchoMRI 3-in-1 Body Composition Analyzer (Echo Medical Systems, Houston, TX, USA) followed by a 48h run in a Comprehensive Lab Animal Monitoring System (CLAMS; Columbus Instruments, Columbus, OH, USA) to measure metabolic parameters and activity (X- and Z-plane). After CLAMS, a glucose tolerance test (GTT) was performed on each female following standard protocols28 (link). Immediately after baseline, females were injected intraperitoneally (i.p.) with glucose (2.0 g/kg body weight) and individually housed in clean cages. After sufficient recovery (~3-4 d), an insulin tolerance test (ITT) was performed after a 5h fast with an i.p. injection of insulin (0.75 units/kg). Blood samples were collected from the tail in individual cages at 15, 30, 60, 90, and 120 min post-injection for both the GTT and ITT. See Supplemental Figure S1 for a graphical illustration of the maternal and adult experimental design.