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Fig. 6 | Journal of Neuroinflammation

Fig. 6

From: Persistent cortical and white matter inflammation after therapeutic hypothermia for ischemia in near-term fetal sheep

Fig. 6

Distribution of cortical microglia and microglial wrapping. A Graph showing the area fraction of NeuN, Iba1 and Hoechst in the cortex of the first parasagittal gyrus in sham control (n = 8), ischemia-normothermia (n = 8) and ischemia-hypothermia (n = 8) groups. *(p < 0.05) vs. sham control, ^(p < 0.05) vs. ischemia-normothermia, one-way ANOVA and LSD post hoc test. B Graph showing the correlation between NeuN area fraction and Iba1 area fraction in the cortex of the first parasagittal gyrus, linear regression. Representative images of Hoechst, Iba1 and NeuN triple-labeling for C sham control with many healthy neurons and few microglia, D ischemia-normothermia with few surviving neurons and an abundance of microglia and E ischemia-hypothermia with many healthy neurons and some microglia. Representative images of maximum intensity projections of z-stacks for Hoechst, Iba1 and NeuN triple labeling for F sham control microglial processes interacting with neurons, G ischemia-normothermia showing the abundance of microglia in close proximity to neurons and H ischemia-hypothermia showing microglial wrapping. Representative images of maximum intensity projections of z-stacks for Hoechst, Iba1, NeuN and TUNEL quadruple labeling for I sham control showing TUNEL negative healthy neurons and ramified microglia, J ischemia-normothermia showing TUNEL-positive neurons wrapped by microglia (arrow with tail) and K ischemia-hypothermia showing TUNEL negative neurons wrapped by microglia (arrow with no tail) and L ischemia-hypothermia showing TUNEL-positive neurons wrapped by microglia (arrow with tail). (1) Hoechst and NeuN, (2) Hoechst and Iba1, (3) Hoechst and TUNEL and (4) all 4 channels merged. Scale bar for H and L4 = 50 µm

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