Fig. 3: Astrocytic deletion of NLG3 impairs hippocampal astrocyte activity.

a, b AAV virus constructs, injection and astrocyte Ca²⁺ imaging in hippocampal slices. c, d Sample traces and summary data of increased GCaMP6f signals in hGFAP-CreERT2 (top; n = 20 cells, 4 mice; p = 0.00203), but not in GFAP-NLG3 KO (bottom; n = 20 cells, 4 mice) astrocytes after HFS. e Reduced activated astrocytes in GFAP-NLG3 KO (n = 10 slices, 4 mice) compared to hGFAP-CreERT2 (n = 11 slices, 4 mice) in response to HFS (p = 0.00000208). f, g Representative traces and summary data of increased GCaMP6f signals in hGFAP-CreERT2 hippocampal astrocytes treated with ACSF (n = 15 cells, 4 mice; p = 0.00732) or AP5 (n = 8 cells, 3 mice; p = 0.0240), but not with PPADS + suramin (n = 15 cells, 4 mice) or MCPG (n = 15 cells, 3 mice) after HFS. h Reduced activated astrocytes in hGFAP-CreERT2 slices treated with PPADS + suramin (n = 6 slices, 3 mice; p < 0.001) and MCPG (n = 5 slices, 3 mice; p < 0.001) compared to those treated with ACSF (n = 6 slices, 3 mice) or AP5 (n = 6 slices, 3 mice) after HFS. i, j AAV virus construct and injections for in vivo fiber photometry Ca²⁺ imaging. k, l Representative traces and summary data of increased astrocytic GCaMP6f signals during social sniffing episodes (pink lines) in GFAP-NLG3 WT (n = 8 recordings, 4 mice; p < 0.001), but not in GFAP-NLG3 KO mice (n = 8 recordings, 4 mice). Data represent mean ± SEM; two-tailed t test for (e); two-tailed paired t test for (d, g); one-way ANOVA with Holm-Sidak test for multiple comparisons for (h), and repeated two-way ANOVA with Holm-Sidak test for multiple comparisons for (l); *p < 0.05, **p < 0.01, ***p < 0.001. Scale bars: 10 μm in left panels of (c); 0.5 dF/F₀ /5 s in right panels of (c, f); 0.2 dF/F₀/60 s in (k). Source data are provided as a Source Data file.