Opposite and Differently Altered Postmortem Changes in H3 and H3K9me3 Patterns in the Rat Frontal Cortex and Hippocampus
Abstract
:1. Introduction
2. Results
3. Discussion
4. Materials and Methods
4.1. Animal Handling and Establishment of Postmortem Delay
4.2. Histology
4.3. Antibodies
4.4. Western Blot Analysis
4.5. Confocal Immunohistochemistry
4.6. Image and Statistical Analyses
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
CNPase | 2′,3′-cyclic nucleotide 3′-phosphodiesterase (EC 3.1.4.37) |
DAPI | 2-[4-(aminoiminomethyl)phenyl]-1H-indole-6-carboximidamide hydrochloride |
GAPDH | glyceraldehyde 3-phosphate dehydrogenase (EC 1.2.1.12) |
GFAP | glial fibrillary acidic protein |
H | histone |
H3K | histone Lys modification |
Iba1 | ionized calcium-binding adaptor molecule 1 |
IgG | immunoglobulin G |
NeuN | neuronal nuclei protein (hexaribonucleotide-binding protein-3 |
PBS | phosphate-buffered saline |
PTM | post-translational modification |
RT | room temperature |
SEM | standard error of the mean |
TBS | Tris-buffered saline |
WB | Western blot |
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Primary Antibody, Abbrev. Name | Primary Antibody, Full Name (Cat. No.) | Final Dilution | Company Name | Secondary Antibody with Fluorochrome, Full Name (Cat. No.) | Company Name | Final Dilution |
---|---|---|---|---|---|---|
NeuN | Mouse anti-NeuN, monocl. ab. (MAB377) | 1:100 | Chemicon, Temecula, CA, USA | Alexa Fluor 488 goat anti-mouse IgG (A-10680) | Invitrogen, Carlsbad, CA, USA | 1:1000 |
GFAP | Mouse anti-GFAP, monocl. ab. (MA1-19395) | 1:100 | ThermoFisher Scientific, Inc., Waltham, MA, USA | Alexa Fluor 488 goat anti-mouse IgG (A-10680) | Invitrogen, Carlsbad, CA, USA | 1:1000 |
CNPase | Mouse anti-CNPase, monocl. ab. (ab6319) | 1:500 | Abcam, Cambridge, UK | Alexa Fluor 488 goat anti-mouse IgG (A-10680) | Invitrogen, Carlsbad, CA, USA | 1:1000 |
Iba1 | Mouse anti-Iba1, monocl. ab. (016-26721) | 1:500 | FUJIFILM Wako Chemicals Europe GmbH, Neuss, Germany | Alexa Fluor 488 goat anti-mouse IgG (A-10680) | Invitrogen, Carlsbad, CA, USA | 1:1000 |
H3 | Rabbit anti-histone H3, polycl. ab., Chip Grade (ab1891) | 1:1500 | Abcam, Cambridge, UK | Alexa Fluor 488 goat anti-rabbit IgG (SAB4600389); Anti-rabbit IgG, perox. conjug. (WB) (A9169) | Sigma, St. Louis, MO, USA; Invitrogen, Carlsbad, CA, USA | 1:2000; 1:1000 (WB) |
H3K9me3 | Rabbit anti-histone H3 (trimethyl K9), polycl. ab., Chip Grade (ab8898) | 1:1000 | Abcam, Cambridge, UK | Alexa Fluor 488 goat anti-rabbit IgG (SAB4600389); Anti-rabbit IgG, peroxidase conjug. (WB) (A9169) | Sigma, St. Louis, MO, USA; Invitrogen, Carlsbad, CA, USA | 1:2000; 1:1000 (WB) |
GAPDH | Mouse anti-GAPDH, monocl. ab., clone GAPDH-71.1 (G8795) | 1:20,000 | Sigma, St. Louis, MO, USA | Anti-mouse IgG, peroxidase conjug. (WB) (A9044) | Sigma, St. Louis, MO, USA | 1:2000 (WB) |
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Dulka, K.; Lajkó, N.; Nacsa, K.; Gulya, K. Opposite and Differently Altered Postmortem Changes in H3 and H3K9me3 Patterns in the Rat Frontal Cortex and Hippocampus. Epigenomes 2024, 8, 11. https://doi.org/10.3390/epigenomes8010011
Dulka K, Lajkó N, Nacsa K, Gulya K. Opposite and Differently Altered Postmortem Changes in H3 and H3K9me3 Patterns in the Rat Frontal Cortex and Hippocampus. Epigenomes. 2024; 8(1):11. https://doi.org/10.3390/epigenomes8010011
Chicago/Turabian StyleDulka, Karolina, Noémi Lajkó, Kálmán Nacsa, and Karoly Gulya. 2024. "Opposite and Differently Altered Postmortem Changes in H3 and H3K9me3 Patterns in the Rat Frontal Cortex and Hippocampus" Epigenomes 8, no. 1: 11. https://doi.org/10.3390/epigenomes8010011