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Fig. 4 | Cellular & Molecular Biology Letters

Fig. 4

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

Fig. 4

In vitro assays confirming the roles of Xbp1 in improving survival and tube formation, and attenuating apoptosis in BMECs. AC MTS assays measured the cell proliferation of brain microvessel endothelial cells (BMECs) after treatment with NC, sh-Xbp1, and ov-Xbp1 for different time intervals, 1 day, 2 days, and 3 days. NC was used as the control group. Proliferation rate = (OD value/(OD value at day 0) − 1) × 100%. Suppression ratio = (1 − experimental OD value/control OD value) × 100%. Results are presented as mean ± SD (n = 3 for each groups). One-way ANOVA followed by post hoc test. *P < 0.05, **P < 0.01 compared with the control. D Representative figures of Transwell (line 1, scale bar 25 μm), tube formation (line 2, scale bar 50 μm; line 3, scale bar 25 μm), and flow cytometry assays (line 4) under NC, ov-Xbp1, and sh-Xbp1 conditions. E Quantitative analysis Transwell assays that showed migration ability of infected BMECs. Results are presented as mean ± SD (n = 3 for each groups). One-way ANOVA followed by post hoc test. *P < 0.05 compared to the control. F The effect of Xbp1 on the formation of tubes in BMECs was explored with the Matrigel assay at magnifications of 40× (tube formation 40) and 100× (tube formation 100). Results are presented as mean ± SD (n = 3 for each groups). One-way ANOVA followed by post hoc test. *P < 0.05, **P < 0.01. G Flow cytometry analyzing apoptosis of BMECs treated with sh-Xbp1 and ov-Xbp1 and control groups. Representative diagrams were shown in the quantification of the early and late apoptotic cells. Results are presented as mean ± SD (n = 3 in each groups). One-way ANOVA followed by post hoc test. *P < 0.05, **P < 0.01 compared with the control. BMEC brain microvessel endothelial cell, FITC fluorescein isothiocyanate, NC normal control, OD optical density, PI propidium iodide, SD standard deviation, Xbp1 X-box binding protein 1

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