Fig. 4

ICV DFX treatment after SAH causes an increase in HO-1/microglia co-localization—Confocal Z-stacked images were taken of the following treatment groups on POD7 in WT mice: SAH sham + vehicle, SAH + vehicle, SAH + IP DFX, and SAH + ICV DFX. a Representative confocal images of each experimental group. All images were taken adjacent to the anterior hippocampus, which can be visualized by the DAPI merged image at the bottom right tile. Each color represents the following: green = HO-1; red = Iba-1/microglia; yellow = overlap of green and red/co-localization of HO-1 and microglia; blue = DAPI nuclei counterstain. All scale bars = 20 μm. b Co-localization heat maps for each experimental group. The x-axis represents channel-1 (HO-1) pixel intensity, while the y-axis represents channel-2 (Iba-1) pixel intensity. Blue represents the lowest population frequency possible while yellow represents the highest. The lower the slope for each heat map, the more HO-1 staining there is per Iba-1/microglia staining. c The percent quantification of co-localization per high-powered field for each experimental group’s confocal Z-stacked images. Co-localization percentage was obtained by dividing the total number of yellow-positive cells by the total number of DAPI-positive cells. The highest HO-1/Iba-1 co-localization percentage was seen in the SAH + ICV DFX group, followed by the SAH + IP DFX and SAH + vehicle group (one-way ANOVA P < 0.05; *P < 0.05; n = 3 per group)