Extended Data Fig. 8: vSub-LH connection is not potentiated after pCFC and inhibition of vSub inputs to LH has no effect on feeding.

a, Representative images showing expression of AAV5-CamkII-ChR2-EYFP in vSub of control and conditioned VGAT-Cre::Ai14 mice from 3 independent experiments. Scale bar, 500 µm. b, Data showing similar fluorescence intensity of ChR2-EYFP expression in vSub of control and conditioned mice. (Mann-Whitney U test, p = 0.743; 8 slices from control and 9 slices from conditioned from n = 3 mice/group) c, Viral injection strategy for ex-vivo whole cell voltage-clamp recording of light evoked synaptic transmission between vSub and ^LHVGAT neurons in VGAT-Cre::Ai14 mice after pCFC paradigm. d, Quantification of light evoked EPSC amplitudes after pCFC in control (9 cells from 3 mice) and conditioned mice (9 cells from 4 mice), each dot represents one neuron (two sided unpaired t-test with Welch’s correction p = 0.6572). e, Data showing similar fluorescence intensity of ChR2-EYFP expression around patched neurons in control (n = 5 neurons) and conditioned (n = 7 neurons) mice (two-sided unpaired t-test with Welch’s correction, p = 0.3692). f, Representative images to show AAV-DIO-hM4D-mCherry expression in ^ArcAgRP, ^TNSST and ^LHVGAT in AgRP-Cre, SST-Cre and VGAT-Cre mice respectively from 3 independent experiments. g, Viral injection strategy for inhibiting vSub inputs to LH. AAVrg-cre virus was injected in LH and cre-dependent AAV-DIO-hM4D-mCherry was injected in vSub. vSub neurons were chemogenetically inhibited before providing food. h, Food intake in saline (black) and CNO (red) injected mice in refeeding after overnight fasting (n = 8, two-sided paired t-test, p = 0.7837). i, Representative image showing expression of AAV-DIO-hM4D-mCherry in vSub from 3 independent experiments. All data in figure are presented as mean values±SEM.