All chemicals and antibodies were obtained from ThermoFisher Scientific, unless otherwise stated. GSPE was obtained from Les Dérives Résiniques et Terpéniques (Dax, France) [16 (link), 17 (link), 19 (link)], and was analyzed in-house, as described in the Supporting Information . GSPE has a total polyphenol content >68%. Results of the analysis are presented in the Supporting Information, Supplementary Fig. 1 and Supplementary Table 1 . Antibodies for HDAC1, HDAC2, HDAC3, acetyl-lysine, acetyl-α-tubulin, acetyl-H3K9, and histone H3 were obtained from Cell Signaling, while those for β-actin and α-tubulin were purchased from Sigma-Aldrich and Santa Cruz Biotech, respectively.
Acetyl h3k9
Manufactured by Cell Signaling Technology
Acetyl-H3K9 is a lab equipment product that detects the acetylation of histone H3 at lysine 9 (H3K9). It is a tool used in epigenetic research to study chromatin modifications and gene regulation.
Automatically generated - may contain errors
Lab products found in correlation
Histone h3, by Cell Signaling Technology (1 mentions)
Acetyl α tubulin, by Cell Signaling Technology (1 mentions)
Acetyl lysine, by Cell Signaling Technology (1 mentions)
Slc7a11, by Abcam (1 mentions)
Vascular endothelial growth factor (vegf), by Cell Signaling Technology (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Sirt6, by Cell Signaling Technology (1 mentions)
Osteocalcin ocn, by Abcam (1 mentions)
Hif 1α, by Cell Signaling Technology (1 mentions)
Runx2, by Cell Signaling Technology (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Nitrocellulose membrane, by Merck Group (1 mentions)
Cox 2, by Cell Signaling Technology (1 mentions)
P ampkα, by Cell Signaling Technology (1 mentions)
Sirt1, by Cell Signaling Technology (1 mentions)
Acetyl h3k14, by Cell Signaling Technology (1 mentions)
P accα, by Cell Signaling Technology (1 mentions)
Ampkα, by Cell Signaling Technology (1 mentions)
Ab16502, by Abcam (1 mentions)
Ab8245, by Abcam (1 mentions)
3 protocols using acetyl h3k9
1
GSPE Modulation of Histone Deacetylases
2
Protein Expression Analysis in Femoral Heads
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Total protein was extracted from the femoral heads and cells of each group with RIPA lysate, and protein concentration was determined with Coomassie brilliant blue. Protein samples of 30 μg were transferred to PVDF (Polyvinylidene Difluoride) membranes after polyacrylamide gel electrophoresis. 5% skimmed milk was blocked for 1 h at room temperature, incubated overnight with primary antibodies for SIRT6 (1 : 1000; cat.no. 12486; Cell Signaling Technology, Inc.), HIF-1α (1 : 1000; cat.no. 14179; Cell Signaling Technology, Inc.), Acetyl-H3K9 (1 : 1000; cat.no. ab10812; Abcam), Runx2 (1 : 1000; cat.no. 12556; Cell Signaling Technology, Inc.), ALP (1 : 1000; cat.no. ab229126; Abcam), osteocalcin (OCN; 1 : 1000; cat.no. ab93876; Abcam), CD31 (1 : 1000; cat.no. 3528; Cell Signaling Technology, Inc.), VEGF (1 : 1000; cat.no. 9698; Cell Signaling Technology, Inc.), transferrin receptor (TFRC; 1 : 1000; cat.no. ab214039; Abcam), divalent metal transporter-1 (DMT1; 1 : 1000; cat.no. ab55735; Abcam), SLC7A11 (1 : 1000; cat.no. ab175186; Abcam), GPX4 (1 : 1000; cat.no. 59735; Cell Signaling Technology, Inc.), and β-actin (1 : 5000; cat.no. AB0035; Abways Technology), and the next day, secondary antibody was added and incubated for 1 h at room temperature for development. β-Actin was used as a reference.
3
Western Blot Analysis of Protein Expression
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Western blot was performed using the protocols described earlier [3] . Equal amounts of protein isolated from pbMEC by RIPA lysis buffer (Beyotime, Shanghai, China) were brie y separated on 10% SDS polyacrylamide gels (GenScript ExpressPlus™ PAGE Gels, Nanjing, China). Then, proteins were transferred onto nitrocellulose membranes (Millipore, Billerica, MA), which were incubated with primary antibodies overnight at 4 °C. After washing for 6 consecutive times, the blots were incubated with horseradish peroxidase-coupled secondary antibody. Differences in protein transfer e ciency between blots were normalised with GAPDH quanti cation. The grey values of the bands of each target protein were quanti ed using the ImageJ system analysis software. Primary antibodies for p-P65, IL1β, TNFα, acetyl-H3K14, acetyl-H3K9, histone H3, p-AMPKα, AMPKα, p-ACCα, ACCα, COX-2 and SIRT1 were purchased from Cell Signaling Technology (Danvers, MA) (#3033, #15101, #3866, #7627, #9649S, #4499, #2535, #5831, #11818, #3676, #12282S, and # 2496S), and p65 was purchased from Abcam (ab16502) and diluted 1:1,000 for incubation. The primary antibody for GAPDH was purchased from Abcam Corporation (ab8245) and diluted 1:5,000 for incubation.
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