Article Figures & Data
Figures
- Figure 1.
Temporal change of AQP4 polarity during the process of pilocarpine-induced TLE. A, Quantitative analysis of AQP4 polarity in control group (CG), 0 d, 1 d, 7 d, 30 d after SE. n = 12, 3 rats per group, 4 slice per rat, ***p < 0.001. B, Representative immunofluorescence images (red, AQP4; blue, DAPI) and gray images of AQP4 at high and low stringency to calculate the AQP4 polarity. Scale bar, 40 μm. 0 d represents the time point immediately after the termination of pilocarpine-induced SE.
- Figure 2.
Effect of the AQP4 polarity distribution inhibitor trifluoperazine on PTZ-induced seizures: A, Seizure latency. B, Seizure duration (*p < 0.05, n = 6 per group). C, Brain edema 3 h after PTZ injection (*p < 0.05, **p < 0.01, n = 3 per group). D, Statistical analysis of hippocampal AQP4 polarity distribution across groups (**p < 0.01, n = 3 animals per group, 2 slices per animal). E, Representative immunofluorescence images: red, AQP4; green, Glut1; blue, DAPI. Scale bar, 40 μm. Trifluoperazine, TFP.
- Figure 3.
Changes in pericyte vascular coverage and PDGF-BB concentration in chronic TLE rats induced by pilocarpine. A, Representative immunofluorescence images of pericyte vascular coverage: red, Pdgfrβ; green, Glut1; blue, DAPI. Scale bar, 50 μm. Yellow arrows indicate pericytes surrounding blood vessels. B, Statistical analysis of pericyte vascular coverage in the hippocampal CA1 region. n = 12, 3 animals per group, 2 brain slices per animal, 2 hippocampal regions. *p < 0.05, **p < 0.01, ***p < 0.001. C, PDGF-BB concentration in serum. n = 8 for control group (CG), 0 d, 1 d, 7 d; n = 6 for 30 d with or without epilepsy. *p < 0.05, **p < 0.01. D, PDGF-BB concentration in CSF. n = 6 for control group, 0 d, 1 d, 30 d with epilepsy; n = 7 for 7 d; n = 5 for 30 d nonepilepsy. *p < 0.05.
- Figure 4.
Effect of pericyte coculture on astrocyte membrane AQP4 levels. HA1800, HBVP, and hCMEC were transwell cocultured with HA1800 cells. AQP4 levels in the membrane of upper layer of HA1800 cell: A, Representative WB images. B, Statistical analysis, n = 3, *p < 0.05. The total AQP4 levels in the upper layer of HA1800 cells: C, representative WB images, D, statistical analysis, n = 3. The uncropped Western blots see Extended Data Figure 4-1.
- Figure 5.
Transcriptomic analysis of lower and upper layer cells after transwell coculture of HA1800, HBVP, hCMEC with HA1800, and HBVP with HBVP cells. Each group contains four specimens. A, Principal component analysis plot. B, Volcano plot of differentially expressed genes (DEG) comparing upper layer HA1800 in HA1800-HBVP to HA1800-HA1800 coculture. C, Volcano plot of DEG comparing upper layer HA1800 in HA1800-hCMEC to HA1800-HA1800 coculture. D, Volcano plot of DEG comparing upper layer HA1800 in HA1800-HBVP to HA1800-hCMEC coculture. E, Volcano plot of DEG comparing lower layer HBVP in HA1800-HBVP to HBVP-HBVP coculture. F–I, GO analysis of DEG in B–E, respectively. The analysis results of AQP4 polarity-related genes associated with GO term “collagen-containing extracellular matrix” is presented in Extended Data Figure 5-1.
- Figure 6.
WB analysis of LAMA1 in lower layer HBVP and hCMEC cells after transwell coculture of HA1800-HBVP, HBVP-HBVP, HA1800-hCMEC, and hCMEC-hCMEC: Representative WB image (A) and statistical analysis (B) of LAMA1 expression, n = 3, **p < 0.01, ***p < 0.001. Changes in hippocampal LAMA1 in chronic TLE rats induced by pilocarpine: representative WB image (C) and statistical analysis (D), n = 3 animals per group, *p < 0.05, **p < 0.01. The uncropped Western blots, see Extended Data Figure 6-1. E, Representative immunofluorescence images of LAMA1 in each group. Red, LAMA1; green, PDGFRβ; blue, DAPI. Yellow arrows indicate LAMA1 localized surrounding pericytes. Scale bar, 50 μm.
- Figure 7.
Analysis of LAMA2 expression in transwell cocultures and hippocampal tissue from pilocarpine-induced chronic TLE rats: A, representative WB image and B, statistical analysis of LAMA2 expression in lower-layer HBVP and hCMEC following transwell coculture of HA1800-HBVP, HBVP-HBVP, HA1800-hCMEC, and hCMEC-hCMEC (n = 3, *p < 0.05, ***p < 0.001). C, Representative WB image and D, statistical analysis of hippocampal LAMA2 expression in chronic epilepsy rats induced by pilocarpine (n = 3 animals per group, *p < 0.05). The uncropped Western blots, see Extended Data Figure 7-1. E, Representative immunofluorescence images of LAMA2 expression in each group. Red, LAMA2; green, PDGFRβ; blue, DAPI. Yellow arrows highlight LAMA2 localization around pericytes.
Extended Data
Figure 4-1
Original Western blots in Figure 4. The quantified bands correspond to the molecular weight indicated in the antibody datasheet of AQP4 and GAPDH. Download Figure 4-1, TIF file.
Figure 5-1
Results of differential expression analysis of AQP4 polarity-related genes associated with the GO term “collagen-containing extracellular matrix”. Download Figure 5-1, XLS file.
Figure 6-1
Original Western blots in Figure 6. The quantified bands correspond to the molecular weight indicated in the antibody datasheet of LAMA1. Download Figure 6-1, TIF file.
Figure 7-1
Original Western blots in Figure 7. The quantified bands correspond to the molecular weight indicated in the antibody datasheet of LAMA2. Download Figure 7-1, TIF file.
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