Immunohistochemistry was performed on formalin-fixed, paraffin embedded mouse and human tissue sections using a biotin-avidin method as described before36 . The following antibodies were utilized: rabbit anti-Amylase (Sigma-Aldrich), pERK1/2 (Cell Signaling), MUC5 (NeoMarkers) and SMYD3 (Abcam). Sections were developed with DAB and counterstained with hematoxylin. Pictures were taken using a Zeiss microscope equipped with the Axiovision software. Analysis of the tumor area and IHC analysis was done using ImageJ software by measuring pixel units.
Smyd3
Manufactured by Abcam
Sourced in United States
SMYD3 is a histone methyltransferase that catalyzes the trimethylation of lysine 4 on histone H3 (H3K4me3). H3K4me3 is a histone modification associated with transcriptionally active chromatin.
Automatically generated - may contain errors
Lab products found in correlation
P erk1 2, by Cell Signaling Technology (2 mentions)
Axiovision software, by Zeiss (2 mentions)
Rabbit anti amylase, by Merck Group (2 mentions)
Protease and phosphatase inhibitor, by Roche (1 mentions)
Detergent compatible protein assay kit, by Bio-Rad (1 mentions)
Nfatc2, by Cell Signaling Technology (1 mentions)
Psmb5, by Cell Signaling Technology (1 mentions)
P ire1α, by Thermo Fisher Scientific (1 mentions)
Isrib, by Meilun (1 mentions)
P perk, by Cell Signaling Technology (1 mentions)
Vcam 1, by Abcam (1 mentions)
Calnexin, by Proteintech (1 mentions)
H3k4me3, by Abcam (1 mentions)
P eif2α, by Cell Signaling Technology (1 mentions)
Gapdh, by Proteintech (1 mentions)
4 protocols using smyd3
1
Immunohistochemical Analysis of Tissue Samples
2
Immunohistochemical Analysis of Tissue Samples
3
Protein Extraction and Immunoblotting
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Cells were lysed by adding Triton X-100 lysis buffer (50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.5% Triton X-100, 5% Glycerol, 1% SDS)–containing protease and phosphatase inhibitors (Roche). The protein quantification was carried out by detergent-compatible protein assay kit (Bio-Rad, Hercules, CA), and 30 μg of protein was resolved in 12% SDS-PAGE gel. The antibodies used in this study are NFATC2, PSMB5, VIM (all from Cell Signaling), ENO3 (Sigma), FOXO1 (Immunoway), SMYD3 (Abcam), FOXP3 (eBioscience), and β-actin (CalBioChem).
4
Unraveling UPR pathway regulation
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Reagent sources were as follows: angiotensin II (Ang II), epigallocatechin gallate (EGCG, a PARP16 inhibitor), 4μ8C (a specific IRE1 inhibitor) and ISRIB (a specific PERK inhibitor) were purchased from Meilunbio (Meilun Biotechnology, Dalian, China). EPZ031686 was purchased from MCE (MedChemExpress, USA). Antibodies were obtained from the following commercial sources: PARP16, Smyd3, H3K4me3, Bip and XBP-1 were purchased from Abcam; VCAM-1, Calnexin, and GAPDH was purchased from Proteintech (Proteintech, USA); IL-6 and iNOS were purchased from Epitomics (Burlingame, CA); p-PERK, p-eIF2α, p53, p21 and p16 were purchased from Cell Signaling Biotechnology (Danvers, MA, USA); p-IRE1α were purchased from Thermo Fisher (MD, USA); ATF6 was purchased from Bioss (Bioss Antibodies, Beijing, China).
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