Extended Data Fig. 6: The PF^Glu neurons project onto ACC^Glu neurons.

a, Schematic of PF injection of AAV-DIO-ChR2-mCherry in CaMKII-Cre mice. b, PF: representative image of mCherry labeling neurons by PF infusion of AAV-DIO-ChR2-mCherry. Scale bar, 200 µm. S2, ZI, SUM, S1HL: images representative of mCherry⁺ fibers in CaMKII-Cre mice with PF injection of AAV-DIO-ChR2-mCherry. Scale bars, 100 µm. c, Schematic of the Cre-dependent retrograde trans-monosynaptic rabies virus tracing strategy in CaMKII-Cre mice. d, Representative images showing input neurons in the S1HL, BLA, DRN, DEn, and IC. Scale bars, 100 µm. e, A series of coronal sections, ipsilateral to site of injection, from a representative mouse showing the major monosynaptic inputs to ACC^Glu neurons. Scale bar, 1 mm. f, Normalized distributions of rabies-labeled input neurons across different brain areas in ipsilateral to the injection site. g, DsRed-labeled neurons expressions at different bregma sites in the thalamus of CaMKII-Cre mice. Scale bar, 200 µm. h, Data showing the number of DsRed-labeled neurons expressions at different bregma sites in the thalamus of CaMKII-Cre mice. S2, secondary somatosensory cortex; ZI, zona incerta; ZID, zona incerta, dorsal part; ZIV, zona incerta, ventral part; SUM, supramammillary nucleus; BLA, basolateral amygdaloid nucleus, anterior part; DRN, dorsal raphe nucleus; DEn, dorsal endopiriform nucleus; IC, insular cortex. All data are presented as the mean ± s.e.m.