Fig. 2

Knockdown of SIRPα enhances efferocytosis by microglia and mitigates neuroinflammation following SAH in vivo. A. Western blot assay for the expression of SIRPα and CD47 in the temporal lobe after SAH. B. Quantification of SIRPα and CD47 protein levels (n = 6 each group). C. RT-qPCR for SIRPα in the temporal lobe after SAH (n = 6 each group). D. Representative immunofluorescence images and three-dimensional imaging showing the extent of efferocytosis following SAH for 24 h. White arrowheads indicate dead/dying neurons that were engulfed by microglia (Neun(red)⁺Iba1(green)⁺). Scale bar, 50 μm. E. Quantification of efferocytosis levels (Neun(red)⁺Iba1(green)⁺ microglia) following SAH for 24 h (n = 6 each group). F. Western blot assay for the expression of IL-1β, IL-18, IL-10, TGF-β1, Bax, Bcl-2 and Cleaved caspase-3 in all groups in the temporal lobe following SAH after knockdown of SIRPα in microglia. G. Quantification of IL-1β, IL-18, IL-10, TGF-β1, Bax, Bcl-2 and Cleaved caspase-3 protein levels (n = 6 each group). H. Skeletonized analysis: Representative images of microglia. Scale bar, 50 μm. I. Quantification of soma area, branch numbers, and total branch length (n = 6 each group). J-K. Representative swimming tracks of the mice in all groups of the MWM tests and quantitative analyses of latency and distance. L. Representative fluorescent images and quantification of TUNEL + neurons following SAH for 24 h. Nuclei were counterstained with DAPI (blue), (n = 6 each group). Scale bar, 100 μm. M. Representative Nissl staining images of the temporal lobe in all groups (n = 6 each group). Scale bar, 50 μm. All values are means ± SEM, ^*P < 0.05, ^**P < 0.01, ^***P < 0.001, ^****P < 0.0001, ns, no significant changes