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Fig. 5 | Cellular & Molecular Biology Letters

Fig. 5

From: X-box binding protein 1 as a key modulator in “healing endothelial cells”, a novel EC phenotype promoting angiogenesis after MCAO

Fig. 5

In vivo assays confirming the roles of Xbp1 in improving survival, attenuating apoptosis, and promoting angiogenesis in BMECs. A Representative images showing the infarct size of ischemic brain dyed with TTC staining. Regions without staining were considered infarct regions. TTC 2,3,5-triphenyl tetrazolium chloride. B Bar plot summarizing the infarct size of ischemic brain under different Xbp1 interventions. Results are presented as mean ± SD (n = 3 in each group). One-way ANOVA followed by post hoc test. *P < 0.05, **P < 0.01 compared to the control. C Representative images showing the survival of endothelial cells by RECA-1 (red) /Hoechst (blue) dual labeling of the brain slices at days 1, 3, 7, and 14 under ov-Xbp1, sh-Xbp1, and NC conditions. Scale bar, 100 μm. D Statistics showing the percentage of RECA-1+ cell area out of each visual fields at days 1, 3, 7, and 14 under ov-Xbp1, sh-Xbp1, and NC conditions. Results are presented as mean ± SD (n = 3 in each groups). Two-way ANOVA. P < 0.05 was considered statistically significant. RECA-1 rat endothelial cell antigen-1, NC normal control. E Representative images showing apoptosis in the brain by TUNEL (green)/DAPI (blue) dual labeling of the brain slices at days 1, 3, 7, and 14 under ov-Xbp1, sh-Xbp1, and NC conditions. Scale bar, 100 μm. F Statistics showing the percentage of the number of TUNEL+ cells out of DAPI+ cells at days 1, 3, 7, and 14 under ov-Xbp1, sh-Xbp1, and NC conditions. Results are presented as mean ± SD (n = 3 in each group). Two-way ANOVA. P < 0.05 was considered statistically significant. TUNEL terminal deoxynucleotidyl transferase (TdT) dUTP nick-end labeling, DAPI 4′,6-diamidino-2-phenylindole. G Representative images showing the proliferating endothelial cells by CD31 (green)/PCNA (red)/DAPI (blue) tri-labeling of the brain slices at days 1, 3, 7, and 14 under ov-Xbp1, sh-Xbp1, and NC conditions. Scale bar, 100 μm. H Statistics showing the percentage of the number of PCNA+/CD31+ dual labeling cells out of CD31+ cells at days 1, 3, 7, and 14 under ov-Xbp1, sh-Xbp1, and NC conditions. Results are presented as mean ± SD (n = 3 in each group). Two-way ANOVA. P < 0.05 was considered statistically significant. PCNA proliferating cell nuclear antigen, DAPI 4′,6-diamidino-2-phenylindole

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