Fig. 6: Hexokinase 2 localizes to mitochondria in microglia.

a Images taken from the cerebral cortex of mitochondria (green) in microglia (magenta) with immunostaining for hexokinase 2 (orange). The images on the right show the boxed region from the low magnification image highlighting the asymmetric distribution of hexokinase 2 within single microglial processes and its mitochondrial association. Arrows indicate processes with low (cyan arrow) and high (white arrow) mitochondrial occupancy. b Images showing mitochondria (green) in microglia (magenta) immunostained for hexokinase 3 (yellow). The images on the right show the boxed region from the low magnification image highlighting the lack of hexokinase 3 labeling and little to no overlap with mitochondria. Arrows indicate processes with low (cyan arrow) and high (white arrow) mitochondrial occupancy. c Area fraction of mitochondria within cytoplasmic ROIs for the microglial processes analyzed in the hexokinase 2 (left) and 3 (right) immunostaining conditions. d Area fraction of hexokinase 2 (left) and 3 (right) within cytoplasmic ROIs of processes with high or low mitochondrial occupancy. e Area fraction of hexokinase 2 versus 3 in mitochondrial ROIs of primary processes with high mitochondrial occupancy. For (c–e), each staining condition had n = 96 primary processes from 24 cells from 8 mice; c, d paired two-tailed t-tests applied; and e unpaired two-tailed t-test with Welch’s correction applied (the line is at the mean ± s.e.m for error bars). For (c–e) see Supplementary Figs. 3f–h and 5g–i for analyses by animal and of sex differences, respectively.