(A) Montage of micrographs of mouse cerebellum labeled for TUNEL (white), GFAP (red), and DAPI (blue) showing TUNEL+ cold injury with surrounding upregulation of GFAP in the cerebellar granule cell layer (dotted lines); boxes show ROIs for ipsilateral injured (INJ) tissue and contralateral control (CTR) tissue; the results shown are representative of 5 mice. (B, C) Micrographs of mouse cerebellum immunolabeled for AQP4 and TRPM4 (B) or SUR1 (C) showing ~3-fold (p=0.004) upregulation of SUR1 and ~3.6-fold (p=0.008) upregulation of TRPM4 in injured AQP4+ cerebellar granule cell layer astrocytes versus control; co-localization analysis showed Pearson’s correlation coefficient of ~0.84 for AQP4 and TRPM4 (B), and ~0.77 for AQP4 and SUR1 (C) in injured tissues; *p<0.05 in t-test versus CTR. (D, E) Immunoblots of co-IP experiments from cold injured and sham injured mouse cerebellum, showing AQP4 immunoblot detecting AQP4 upon TRPM4 immunoprecipitation (IP) in injured tissues with low (low ex) and higher exposures (high ex) (D), and SUR1 immunoblot detecting SUR1 upon AQP4 IP in injured tissues (E); the results shown are representative of 3 replicates.