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5 protocols using xav 939 s1180

1

Protein Expression and Apoptosis Assay

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Antibodies for UBE2S (14115–1-AP) and β-actin (66009–1-Ig) were purchased from Proteintech (Wuhan, China). Antibodies for PARP (9532) and caspase-3 (9662) were obtained from Cell Signaling Technology (Danvers, MA, USA). Antibodies for c-Myc (ab32072), β-catenin (ab32572), and cyclin D1 (ab16663) were acquired from Abcam (Cambridge, UK). XAV-939 (S1180) and Olaparib (S1060) were obtained from Selleck Chemicals (Houston, TX, USA). Tosyl Arginine Methyl Ester (TAME, 172104) was purchased from Sigma-Aldrich (St. Louis, MO, USA).
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2

Investigating SHMT2 and BRCC36 Regulation

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The siRNA oligos targeting SHMT2 and BRCC36 were obtained from Ruibo (Shanghai, China). The target sequences of the siRNA oligos were as follows.
si-SHMT2 #1, 5' CCACAATCATGCCATTGCT3'
si-SHMT2 #2, 5'CCTGCAGGTTCTGAAGAAT3'
si-BRCC36 #1, 5'GCAGGAATTACAACAAGAA3'
si-BRCC36 #2, 5'GAAAGTGTGCCTTGAATCA3'
Proteasome inhibitor MG132 (S2619), Wnt pathway inhibitors XAV939 (S1180), and iCRT-14 (S8704) were purchased from Selleck (TX, USA). Cycloheximide (CHX) (239763) was purchased from Millipore (Darmstadt, Germany).
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3

Quantitative Analysis of Wnt Signaling Pathway

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Anti-LXRα (ab41902), anti-β-catenin (ab32572), anti-CD44 (ab51037), and anti- Cyclin D1 (ab134175) antibodies were supplied by Abcam (Abcam, USA). GAPDH (10949-1-AP) and β-tubulin (10094-1-AP) antibodies were supplied by Proteintech Group (Proteintech, USA). Dimethyl sulfoxide and GW3965 were purchased from Sigma-Aldrich (St Louis, MO, USA). XAV939 (S1180) and Wnt agonist 1 (S8178) were purchased from Selleck (Selleck, USA).
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4

Wnt/β-catenin Signaling Pathway Analysis

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Antibodies directed against β-catenin, p-β-catenin, E-cadherin, N-cadherin, Bax, Bcl, c-caspase 3, caspase 3, β-actin, and β-tubulin were purchased from Cell Signaling Technologies (Danvers, MA, USA). Antibodies directed against histone H3 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies directed against WDR74 were purchased from OriGene (Rockville, MD, USA). Antibodies directed against lef1, TCF1, c-myc, and cyclin D1 were purchased from Abisin (Shanghai, China). XAV-939(S1180), a Wnt/β-catenin signaling pathway-specific inhibitor, was purchased from Selleck Chemicals (Houston, TX, USA).
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5

TRPA1 Signaling Pathway Regulation

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TRPA1 (19124-1-AP) was purchased from Proteintech Group (Rosemont, IL 60018, USA). CaMKII (4436), pCaMKII (12716), β-catenin (8480) were purchased from Cell Signaling Technology (Danvers, Massachusetts, USA). Retinal (R2500), JT010 (SML1672), HC030031 (H4415) were purchased from Sigma Aldrich (Darmstadt, Germany). Dimethyl sulfoxide (DMSO), RIPA buffer (R0100), BCA protein assay kit (PC0020), poloxamer gel (Polyethylene-polypropylene glycol 407, S7071) and formalin (G2161) were purchased from Solarbio (Beijing, China). Protease inhibitor cocktail (B14011) was purchased from Bimake (Shanghai, China). XAV-939 (S1180) was purchased from Selleck Chemicals (Shanghai, China). Fluo-4 AM (ab142773), DIO Staining Solution (C1038) were purchased from Beyotime (Shanghai, China). Stock solutions were prepared as follows: HC030031, and XAV-939 were dissolved in DMSO, JT010, and retinal in absolute ethanol. Masson-Fontana staining solution (G2032) was purchased from Solarbio (Beijing, China). Opti-MEM medium (31985062), Lipofectamine 2000 (11668030) and chemiluminescence (ECL) were obtained from Thermo Fisher Scientific (Massachusetts, USA). The UVA or UVB source used was a 9W UVA or UVB broadband lamp (Philips, Eindhoven, Netherlands) and radiation energies were measured using a UVX radiometer (UVP, Upland, California, USA).
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