Journal: bioRxiv

Article Title: Persistent DNA methylation and downregulation of homeostatic genes in astrocytes after pilocarpine-induced status epilepticus: Implications for epileptogenesis

doi: 10.1101/2024.12.31.630767

Figure Lengend Snippet: A- B-C : Representative images of 5mC/GFAP immunostaining in hippocampal brain sections of animals exposed to pilocarpine-induced SE at 7 days post-SE (DPSE) (A), 21DPSE (B) or 35DPSE (C). D : High magnification images showing details of the CA-1 area and astrocytes immunolabeled by 5mC at 35DPSE. E : Quantitative analysis of the 5mC abundance in GFAP+ cells as mean fluorescence intensity (MFI). Data are presented as mean ± SEM and significance was analyzed by two-way ANOVA and Tukey post-test. N= 5-7 animals per group.

Article Snippet: Primary antibodies from different brands were used as follows: rabbit polyclonal anti- glutamine synthetase (ThermoFisher, RRID:AB_2546416; 1/500), chicken polyclonal anti- Glial Fibrillary Acidic Protein (GFAP) (ThermoFisher, RRID:AB_1074620; 1/500), mouse monoclonal anti-GFAP (Sigma, RRID:AB_477010; 1/1000), rabbit polyclonal anti-GFAP (Dako, RRID:AB_10013382; 1:3000), rabbit polyclonal anti-AQP4 (Santa Cruz, RRID:AB_2274338; 1/800); mouse monoclonal anti-5-methylcytosine (5mC) (Zymo Research cat# A-3001, 1/1000).

Techniques: Immunostaining, Immunolabeling, Fluorescence

Journal: bioRxiv

Article Title: Persistent DNA methylation and downregulation of homeostatic genes in astrocytes after pilocarpine-induced status epilepticus: Implications for epileptogenesis

doi: 10.1101/2024.12.31.630767

Figure Lengend Snippet: A : Astroglial cultures were exposed to HMGB1 500 ng/ml for 18 h and allowed to recover in the control tissue culture medium for 0, 24 h, 72 h, or 7 days. Representative images of the 5mC/GFAP immunostaining after the indicated times. The graph shows the quantitative analysis of the mean fluorescence intensity (MFI) in GFAP-immunoreactive cells in the culture relative to the control conditions. Results are presented as mean ± SE and statistical analysis was performed with two-way ANOVA and Tukey post-test. N= 9. B : The heat map illustrates the gene expression levels analyzed by real time-RT-qPCR at the different recovery time points after HMGB1 exposure. Data were log2-transformed after applying the ΔΔCt method to facilitate comparison. C : Detailed relative mRNA levels of the results presented in the heat map with the statistical analysis. Results are presented as mean ± SE and statistical analysis was performed with two-way ANOVA and Tukey post-test. N= 6.

Article Snippet: Primary antibodies from different brands were used as follows: rabbit polyclonal anti- glutamine synthetase (ThermoFisher, RRID:AB_2546416; 1/500), chicken polyclonal anti- Glial Fibrillary Acidic Protein (GFAP) (ThermoFisher, RRID:AB_1074620; 1/500), mouse monoclonal anti-GFAP (Sigma, RRID:AB_477010; 1/1000), rabbit polyclonal anti-GFAP (Dako, RRID:AB_10013382; 1:3000), rabbit polyclonal anti-AQP4 (Santa Cruz, RRID:AB_2274338; 1/800); mouse monoclonal anti-5-methylcytosine (5mC) (Zymo Research cat# A-3001, 1/1000).

Techniques: Control, Immunostaining, Fluorescence, Expressing, Quantitative RT-PCR, Transformation Assay, Comparison

Journal: bioRxiv

Article Title: Persistent DNA methylation and downregulation of homeostatic genes in astrocytes after pilocarpine-induced status epilepticus: Implications for epileptogenesis

doi: 10.1101/2024.12.31.630767

Figure Lengend Snippet: A : Double immunostained hippocampal brain sections form resected foci in TLE patients studied with anti-GFAP (green) and anti-5mC (red). B : Sections from post-mortem control material showed that the same hippocampal regions for GFAP/5mC showing a reduced degree of methylation. C : Quantitative analysis of the assay was presented as mean ± SEM and statistical analysis was performed with an unpaired t-test. N = 4 (TLE); N= 3 (controls).

Article Snippet: Primary antibodies from different brands were used as follows: rabbit polyclonal anti- glutamine synthetase (ThermoFisher, RRID:AB_2546416; 1/500), chicken polyclonal anti- Glial Fibrillary Acidic Protein (GFAP) (ThermoFisher, RRID:AB_1074620; 1/500), mouse monoclonal anti-GFAP (Sigma, RRID:AB_477010; 1/1000), rabbit polyclonal anti-GFAP (Dako, RRID:AB_10013382; 1:3000), rabbit polyclonal anti-AQP4 (Santa Cruz, RRID:AB_2274338; 1/800); mouse monoclonal anti-5-methylcytosine (5mC) (Zymo Research cat# A-3001, 1/1000).

Techniques: Control, Methylation

Journal: bioRxiv

Article Title: Persistent DNA methylation and downregulation of homeostatic genes in astrocytes after pilocarpine-induced status epilepticus: Implications for epileptogenesis

doi: 10.1101/2024.12.31.630767

Figure Lengend Snippet: A : Double immunostained hippocampal brain sections form resected foci in TLE patients or control hippocampal brain sections studied with anti-GFAP (green) and anti-Kir4.1 (red). B : Quantitative analysis of the assay was presented as mean ± SEM and statistical analysis was performed with an unpaired t-test. N = 4 (TLE); N= 3 (controls). C : Double immunostained hippocampal brain sections form resected foci in TLE patients or control hippocampal brain sections studied with anti-GFAP (green) and anti-GS (red). D : Quantitative analysis of the assay was presented as mean ± SEM and statistical analysis was performed with an unpaired t-test. N = 4 (TLE); N= 3 (controls).

Article Snippet: Primary antibodies from different brands were used as follows: rabbit polyclonal anti- glutamine synthetase (ThermoFisher, RRID:AB_2546416; 1/500), chicken polyclonal anti- Glial Fibrillary Acidic Protein (GFAP) (ThermoFisher, RRID:AB_1074620; 1/500), mouse monoclonal anti-GFAP (Sigma, RRID:AB_477010; 1/1000), rabbit polyclonal anti-GFAP (Dako, RRID:AB_10013382; 1:3000), rabbit polyclonal anti-AQP4 (Santa Cruz, RRID:AB_2274338; 1/800); mouse monoclonal anti-5-methylcytosine (5mC) (Zymo Research cat# A-3001, 1/1000).

Techniques: Control