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. 2020 May 26;9:e53074. doi: 10.7554/eLife.53074

Figure 2. WT:AZIP-Chimeric mice in b6 background have normalized adiposity and fully complemented metabolic parameters on chow diet.

(A) Body weight of AZIPs (solid red), WT:AZIP-chimeras (open red), and WT:WT-chimeras (black triangle) from 3 weeks to 24 weeks old. No difference in body weight was observed between WT:AZIP- and WT:WT-chimeras at all ages measured, except for AZIP mice that show significantly lowered body weight at weaning age (3 and 4 weeks old) but no difference is detected afterwards. (B) Body fat content measured by MRI at 12 weeks versus chimerism. There were no significant differences between AZIP and WT:AZIP-chimeric mice but WT:AZIP-chimeric mice are generally leaner than WT:WT-chimeric mice. (C) Serum leptin levels versus chimerism. AZIP mice have non-detectable levels of leptin and WT:AZIP-chimeras show serum leptin levels ranging from 0.6 to 12.5 ng/mL that is intermediate to AZIPs and WT:WT chimeras. No correlation is seen between chimerism and leptin levels in WT:AZIPs and WT:WTs. (D) Serum leptin and percent fat content correlate well in both WT:AZIP- and WT:WT-chimeras. There is no significant difference between the slopes of the two linear regressions. Two statistical outliers of WT-chimera with high leptin were excluded. Together, the results show that AZIP-chimeras form adipose tissue with normal leptin production despite a general reduction in adiposity. (E) Blood glucose levels at 12 weeks old. AZIPs are significantly higher than WT:AZIP and WT:WT-chimeras (297 ± 24 mg/dL for AZIPs vs. 193 ± 11 mg/dL for WT:AZIPs p=0.0005) and WT:AZIPs and WT:WT were the same (193 ± 11 mg/dL for WT:AZIPs vs 182 ± 7 mg/dL for WT:WTs, p=0.4). (F) Insulin level at 12 weeks. AZIP hyperinsulinemia was rescued in WT:AZIP-chimeras, from 117 ± 10 ng/mL to 8.2 ± 3.6 ng/mL (p<0.0001). (G) Liver weight. Liver weight is normalized by complementation (5.8 ± 0.5 for AZIPs vs 2.1 ± 0.2 g in WT:AZIPs; p=6xE-4 and 1.7 ± 0.1 g for WT:WTs). For (A) to (F), n = 17 for AZIP, n = 17 for WT:AZIP, and n = 19 for WT:WT-chimeras. For (G), n = 9 for AZIP, n = 12 for WT:AZIP and n = 14 for WT-chimeras, and liver analysis are performed at 6 months old.

Figure 2.

Figure 2—figure supplement 1. B6 Chimeras on Chow Diet.

Figure 2—figure supplement 1.

(A) Body fat content measured by MRI at 12 weeks. There were no significant difference between AZIP and WT:AZIPs and WT:AZIP-chimeric mice are generally leaner than WT:WT-chimeric mice. (B) Serum leptin levels of three groups of animals. Complementation by ES-cells rescued hypoleptinemia phenotype, where non-chimeric AZIP mice have non-detectable levels of leptin and AZIP-chimeras show serum leptin levels ranging from 0.6 to 12.5 ng/mL. (C) Microscopy study of epididymal white adipose tissue (visceral) and inguinal subcutaneous white adipose tissue (SubQ) of WT:AZIP- and WT:WT-chimeras. AZIP-chimera shown here is 10% chimeric and WT-chimera was 8% chimeric. Brown color represents staining for YFP. Note comparable cell size in both tissues. (D) Chimerism versus blood glucose in WT:AZIP- and WT:WT-chimeras. No difference was detected between high and low percent chimerism. (E) Liver wet weight in mg in WT:AZIP and WT:WT chimeras show no relationship to degree of chimerism. For (A), (B) and (D), n = 17 for AZIP, n = 17 for WT:AZIP-chimeras, and n = 19 for WT:WT-chimeras. For (E), n = 12 for WT:AZIP-chimera, and n = 14 for WT:WT-chimeras, and liver analysis are performed at 6 months old.