Extended Data Fig. 5: Functional expression of hM3Dq in Hes5⁺ SDH astrocytes.

a, Schematic illustration of the experimental protocol and timeline in Hes5-CreERT2 mice using the Cre-On system. AAV-hM3Dq^FLEX was microinjected unilaterally into the SDH of Hes5-CreERT2 mice. Tamoxifen treatment induced the expression of hM3Dq in Cre-positive (Hes5⁺) cells. b, Double immunostaining of hM3Dq (HA-tag) and cell-type markers (NeuN, IBA1 and APC) in the SDH of Hes5-CreERT2;AAV-hM3Dq^FLEX mice. The data are representative of three independent experiments. c, Representative images of the Ca²⁺ response (left) and an example of traces and average data (right) for the effect of CNO treatment on the fluorescence intensity of ROIs in wild-type and Hes5-CreERT2;AAV-hM3Dq^FLEX mice, both of which expressed GCaMP6m in SDH astrocytes. CNO induced astrocytic Ca²⁺ increases in spinal cord slices taken from Hes5-CreERT2;AAV-hM3Dq^FLEX mice but not from wild-type;AAV-hM3Dq^FLEX mice. The data are representative of three independent experiments. Scale bars, 20 μm (b) and 100 μm (c).