Fig. 1: Sphk1 expression is significantly increased in peri-hematomal endothelial cells after intracerebral hemorrhage (ICH).

A RT-qPCR analysis of peri-hematomal SPHK1 mRNA expression levels in patients with ICH (n = 6 patients/group). B, C Western blotting analysis of peri-hematomal Sphk1 protein levels in patients with ICH (n = 6 patients/group). D ELISA analysis of patient serum S1P levels (n = 6 patients/group). E Schematic for the generation and analysis of the mouse ICH model. F ELISA analysis of mouse serum S1P levels at 24 h after ICH (n = 6 mice/group). G RT-qPCR analysis of peri-hematomal Sphk1 mRNA expression levels at 24 h after ICH in mice (n = 6 mice/group). H, I Western blotting analysis of peri-hematomal Sphk1 protein levels at 24 h after ICH in mice (n = 6 mice/group). J RT-qPCR analysis of the time-course changes of Sphk1 mRNA levels after ICH in mice (n = 3 mice/time point, comparisons were conducted between the Sham and ICH mice at different time points). K, L Western blotting analysis of the time-course changes of Sphk1 protein levels after ICH in mice (n = 4 mice/time point, comparisons were conducted between the Sham and ICH mice at different time points). M Immunofluorescence staining of peri-hematomal Sphk1 with the astrocyte marker GFAP, neuron marker NeuN and endothelial cell marker CD31 in brain tissue from the sham mice and the ICH mice at 24 h after surgery. Scale bar=20 μm. N Quantitative analysis of Sphk1 fluorescence signal density after ICH in mice (n = 6 mice/group). Sphk1 fluorescence signal co-localized with respective cellular marker was quantified and normalized by the cellular marker area. Data are expressed as means ± SEM; ns p > 0.05, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, two-tailed unpaired Student’s t-test except for J and (L), which used one-way ANOVA and Tukey multiple comparisons test.