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. 2023 Jan 24;60:102615. doi: 10.1016/j.redox.2023.102615

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© 2023 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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Fig. 4 — Activation of SIRT1 protects the BSCB and promotes functional recovery after SCI. Vehicle or SRT1720 (50 μg/kg) was intrathecally injected into wild-type C57BL/6J mice 1 h after SCI and continued for 3 consecutive days. (A) Representative images of spinal cords, showing EB extravasation at 3 d after SCI in mice treated with vehicle or SRT1720 (n = 6 animals per group). The spinal cords from the mice treated with SRT1720, compared with vehicle, showed significantly less EB extravasation 3 d after SCI. (B) Immunofluorescence images of EB extravasation 1 mm caudal to the lesion epicenter in the transverse sections of the spinal cord at 3 d after SCI in mice treated with vehicle or SRT1720 (n = 6 animals per group). (C) Quantification of EB fluorescence intensity for each group (n = 6 animals per group). (D) Quantification of EB content by spectrophotometry in the spinal cord at 3 d after SCI in mice treated with vehicle or SRT1720 (n = 6 animals per group). (E) Representative images of homogenized spinal cords, showing tissue blood at 3 d after SCI in mice treated with vehicle or SRT1720 (n = 6 animals per group). (F) Representative immunofluorescence images of ZO-1 (green) and CD31 (red) in the spinal cord at 3 d after SCI in mice treated with vehicle or SRT1720 (n = 6 animals per group). Images of selected regions are shown at higher magnification. The sections from mice treated with SRT1720 compared with vehicle control showed significantly greater endothelial ZO-1 expression at 3 d after SCI. (G) Quantification of endothelial ZO-1 expression (n = 6 animals per group). (H) Representative immunoblots of the TJ proteins ZO-1, occludin and claudin5 in the spinal cord at 3 d after SCI in mice treated with vehicle or SRT1720 (n = 6 animals per group). (I) Representative TEM images of TJ ultrastructure in the spinal cord at 3 d after SCI in mice treated with vehicle or SRT1720. Images of selected regions are shown at higher magnification. Red squares and red arrows indicate the TJs (n = 6 animals per group). (J, K) Quantification of the width and length of TJs (n = 6 animals per group). (L) Representative immunofluorescence images of NeuN (green) and F4/80 (red) in the spinal cord at 7 d after SCI in mice treated with vehicle or SRT1720 (n = 6 animals per group). The sections from the mice treated with SRT1720, compared with vehicle control, showed significantly less inflammatory cell infiltration and more neurons at 7 d after SCI. (M) Quantification of the area of F4/80⁺ inflammatory cells (n = 6 animals per group). (N) Quantification of the number of NeuN⁺ neurons (n = 6 animals per group). (O) BMS scores during 28 days of recovery after SCI in mice treated with vehicle or SRT1720 (n = 12 animals per group). The mice treated with SRT1720, compared with vehicle control, showed better behavioral recovery over the course of 28 days after SCI. (P) Representative images of footprint analysis at 28 d after SCI in mice treated with vehicle or SRT1720 (n = 12 animals per group). (Q, R) Quantification analysis of stride length and width (n = 12 animals per group). *P < 0.05 compared with the vehicle control group. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)