Skip to main content
. Author manuscript; available in PMC: 2018 Oct 18.
Published in final edited form as: Nature. 2018 Apr 18;556(7702):505–509. doi: 10.1038/s41586-018-0049-7

Figure 2. CRISPR-mediated deletion of leptin receptors in AgRP neurons results in severe obesity and diabetes.

Figure 2

a, b, Schematic diagram of AAV pU6-sgRNALepr::pEF1α-FLEX-mCherry (AAV-sgLepR) injected bilaterally into the ARC of Agrp-IRES-Cre::LSL-Cas9-GFP mice and mCherry::GFP co-immunostaining. c, d, Coimmunostaining of mCherry and leptin-induced phosphorylated STAT3 (Tyr705, pSTAT3) in virus-transduced Agrp-IRES-Cre (Cas9−, upper panel) and Agrp-IRES-Cre::LSL-Cas9-GFP mice (Cas9+, lower panel) (n=4 mice per group). e–l, Representative littermates (e), serum leptin levels (f), body weight and analysis of fat mass (insert) (g), daily food intake (h), oxygen consumption (i), ad libitum fed blood glucose levels (j), Glucose-tolerance test (k), and Insulin-tolerance test (l) at 8 weeks post-viral injection (n=9 mice per group). m, Growth curve following viral injection into the ARC of 4-week-old Agrp-IRES-Cre (Cas9−), Agrp-IRES-Cre::LSL-Cas9-GFP (Cas9+), Leprdb/+ (db/+), and Leprdb/db (db/db) mice (n=9 mice per group). n, o, Schematic diagram and blood glucose measurement in STZ-treated, virus-transduced Agrp-IRES-Cre (Cas9−) and Agrp-IRES-Cre::LSL-Cas9-GFP mice (Cas9+) mice following chronic administration of saline/leptin into the lateral ventricle with implanted osmotic pump (n=9 mice per group). Data are mean ± s.e.m. and representative of three independent experiments; *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001; Student’s two-tailed, unpaired t-test (d, f, g-insert, h, j) or two-way ANOVA (g, k, l, m, o) with Šidák post hoc test (k).