Fig. 3: Comparison between SIMPLeR and SIMPLE.

a–d Less MNPs in the SIMPLeR territory than in the SIMPLE-induced hypoperfusion in the dMCA territory. a, b Representative LSCI of two live P3 mouse pups at 0.5 h of SIMPLeR and SIMPLE using the same MNP dose (6 mg kg⁻¹). (c) LSCI images of a P5 mouse pup insulted by 0.5 h of SIMPLE using 80 mg kg⁻¹ MNPs. d Quantification of the time course of cerebral blood flow change in the ischemic hemisphere normalized to that of the contralateral side for a, b, and c (N = 4 mice for each time point). For each time point, the effect sizes of SIMPLeR versus SIMPLE (6 mg kg⁻¹) are 12.9, 26.4, 18.5, and 38.4 and those of SIMPLE 80 mg kg⁻¹ versus 6 mg kg⁻¹ are 15.8, 42, 19.7, and 39.2. e–g Representative images of Fluoro-Jade C staining of brain sections from P3 mouse pups insulted with SIMPLeR and SIMPLE. h Quantification of the volume of Fluoro-Jade C -positive brain regions for e, f, and g (N = 4 mice per group). The effect sizes of SIMPLeR and SIMPLE (80 mg kg⁻¹) relative to the reference group SIMPLE (6 mg kg⁻¹) are 34.3 and 60.3, respectively. (i) Macroscopic images of TTC staining of the whole brain (P6) and brain sections (P4) from mouse pups after 10 h of focal ischemia. T2W MRI of P3 mouse pups at 7 h (j) and P5 at 14.5 h (k) after SIMPLE. l Quantification of infarcted brain volume for j and k. m The second MRI T2W image for the same mouse in j at P50. Pink and green dashed lines indicate ischemic ipsilateral and contralateral cerebral cortex, respectively. n Stitched TEM images of the dMCA segment shown in Fig. 2f, g, and Supplementary Fig. 5. Magenta arrows show MNPs bound to RBCs; vascular wall layers are marked in magenta and green. o Stitched TEM image of a red embolus located at the dMCA of P3 mouse pups that underwent 6-hour SIMPLeR. RBCs are marked in red, and surrounding fibrin is marked in blue. Blue line outlines the edge of fibrin in Insert. p–r Stitched TEM images of the dMCA segment (Supplementary Fig. 6) of P3 mouse pups with SIMPLE. Aggregated MNPs (180 nm) are outlined by green lines in p. The enriched white blood cells and platelets adjacent to the MNP occlusion are shown in q and r, respectively. s-s-ii Statistical analysis of different components of white thrombus-like obstructions or red emboli for n–r. A total of 766 μm² (n), 319 μm² (o) and 537 μm² (p, q, r) TEM images were analyzed. *p < 0.05, ****p < 0.0001, ns not significant. Data are presented as Mean ± SEM.