Extended Data Fig. 9: In-vitro and in-vivo validation of AAV-DIO-miR-OxR2 virus.
From: Orexin signaling in GABAergic lateral habenula neurons modulates aggressive behavior in male mice
a, N2A cells treated with miR-OxR2 construct selectively reduced OxR2 expression compared to cells treated with miR-scrambled construct, but did not reduce expression of related transcripts (two-tailed student’s t-test, n = 3 biologically independent plates per group, 3 replicates per plate; OxR2: t(4)=2.402, p = 0.0482; OxR1: t(4)=0.2123, p = 0.8423; Avpr2: t(4)=0.3686, p = 0.7311; Htr3a: t(4)=1.309, p = 0.2607; Drd4: t(4)=0.1925, p = 0.8567; Nmur: t(4)=1.672, p = 0.1699; Drd1: t(4)=0.9239, p = 0.4078; Mchr1: t(4)=1.467, p = 0.2163; Glpr1: t(4)=1.785, p = 0.1488). b, GAD2-Cre mice injected with AAV-DIO-miR-OxR2 displayed reduced expression of OxR2 compared to mice injected with AAV-DIO-miR-scrambled as determined by ISH (two-tailed student’s t-test, n = 3 mice, 2 slices per mouse, t(4)=18.44, p = 0.0001). c, Representative image of GFP expression localized to GAD2 LHb neurons in GAD2-Cre mice injected with AAV-DIO-miR-OxR2, scale bar = 25 μm. Experimental images were obtained from 6 biologically independent mice, 2 slices per mouse, with similar results obtained. d, Representative images of OxR2 expression in GAD2 LHb neurons infected with AAV-DIO-miR-OxR2 or AAV-DIO-miR-scrambled, scale bar, 20 μm. *p < 0.05, ***p < 0.001. All data are expressed as mean ± SEM.
