Extended Data Fig. 9: Characterization of tamoxifen-inducible astrocytic α7-nAchR conditional knockout mice (hGFAP-CreER^T2/Chrna7^loxp/loxp, cKO).

a, PCR analysis of hGFAP-CreER^T2:Chrna7^WT/WT (cWT) and hGFAP-CreER^T2:Chrna7^loxp/loxp mice (α7-cKO mice). The experiment was repeated before immunostaining and two-photon imaging experiment. b, c, α7-nAchR immunostaining in the auditory cortex of a cWT (b) or a cKO (c) mouse indicates a pronounced loss of α7-nAchR-positive astrocytes in cKO. Left, neuronal staining (neuronal marker: NeuN) in L1 and L2/3 in the auditory cortex. Right, astrocyte staining (astrocyte marker: S100β) in the auditory cortex. One astrocyte from panel b was shown in Fig. 7b in an expanded scale. d, Bar graphs summarizing the α7-nAchR expression in neurons in the auditory cortex of cWT and cKO mice. n = 12 sections from 4 mice in each group (cWT versus cKO, Z = −0.6067, P = 0.5440, two-sided Wilcoxon rank-sum test). e, Bar graphs summarizing the α7-nAchR expression in astrocytes in the auditory cortex of cWT and cKO mice. n = 16 sections from 5 mice in cWT group; n = 11 sections from 4 mice in cKO group. (cWT versus cKO, Z = 4.3257, P = 1.52 E-5, ***P < 0.001, two-sided Wilcoxon rank-sum test). f, In vivo two-photon Ca²⁺ imaging of L1 interneurons in the auditory cortex of a cKO mouse, with dashed lines indicating neurons of interest. g, Single trials and their average of Ca²⁺ responses to footshock from these neurons as indicated in panel f. h, Bar graph summarizing the fraction of L1 neurons that responded to footshock in cWT and cKO mice. n = 9 fields of view, 42 cells from 3 mice in cWT group; n = 15 fields of view, 127 cells from 6 mice in cKO group (cWT versus cKO, Z = 0.2684, P = 0.7884; two-sided Wilcoxon rank-sum test). All data in the figure are shown as mean ± s.e.m.

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