Fig. 5

Otp expression in AgRP-neurons and expression of arcuate neuropeptides in Otp-null embryos. a IHC analyses of Otp expression in Npy-hrGfp embryos using our home-made antibodies against Otp. 86 ± 3% of GFP⁺ cells were Otp⁺ cells in IHC. b A confocal Z-stack analysis was also performed with IHC analyses of Otp expression in E16.6 Npy-hrGfp embryos. c ISH for GHRH combined with IHC for Otp. 3 ± 1% of GHRH⁺ cells were Otp⁺ cells in IHC. d ISH analyses of E16.5 Otp-null embryos (n = 3) revealed a significant decrease in the expression of NPY and Sst but neither GHRH nor POMC in comparison to their littermate controls (n = 4). e IHC analyses of E16.5 Otp-null embryos (n = 4) revealed a significant decrease in the number of Otp⁺, GR⁺, and Bsx⁺ cells but neither Dlx1⁺ or TH⁺ cells relative to their littermate controls (n = 4). Quantification was performed with multiple embryos as indicated and at least three sections from each embryo, and representative images for only one side of the ARC are as shown. Scale bars, 100 μm. Student’s t-test results are as indicated: ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗∗p < 0.0001 (d, e). Bars represent mean, error bars represent the SEM