Total RNA was extracted from tissues and cells by using Trizol reagent (Invitrogen) according to the manufacturer’s instructions. Total RNA was reverse-transcribed by using a PrimeScript RT reagent kit (Takara Bio Inc.), and small RNA polyadenylation was performed (Zhang et al., 2012 (link)). Real-time PCR was performed on a real-time PCR system (ABI 7900; Applied Biosystems). The primer sequences are provided in Table S3 . Mouse monoclonal anti–TEF-1 (610922; BD), rabbit polyclonal anti-MyoD (sc-760; Santa Cruz Biotechnology, Inc.), rabbit polyclonal anti-MEF2 (sc-313; Santa Cruz Biotechnology, Inc.), mouse monoclonal anti-myogenin (sc-12732; Santa Cruz Biotechnology, Inc.), mouse monoclonal anti–α-tubulin (T6199; Sigma-Aldrich), mouse monoclonal anti–β-actin (A5316; Sigma-Aldrich), mouse monoclonal anti–glyceraldehyde 3-phosphate dehydrogenase (anti-GAPDH; KC-5G4; KangChen Bio-tech Inc.), rabbit polyclonal anti-TEAD2 (21159-1-AP [Proteintech]; 33900 [Signalway Antibody]), rabbit polyclonal anti-TEAD3 (13120-1-AP; Proteintech), and rabbit polyclonal anti-TEAD4 (12418-1-AP) antibodies were used for Western blot analysis.
Sc 313
Manufactured by Santa Cruz Biotechnology
The sc-313 is a laboratory instrument designed for use in scientific research. It is a product of Santa Cruz Biotechnology. The core function of the sc-313 is to perform specific tasks related to biological and chemical analysis, as required by researchers and scientists.
Automatically generated - may contain errors
Lab products found in correlation
Abi 7900, by Thermo Fisher Scientific (1 mentions)
Mouse monoclonal anti α tubulin, by Merck Group (1 mentions)
Mouse monoclonal anti β actin a5316, by Merck Group (1 mentions)
Anti myod sc 760, by Santa Cruz Biotechnology (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Primescript rt reagent kit, by Takara Bio (1 mentions)
β actin, by Merck Group (1 mentions)
P creb, by Santa Cruz Biotechnology (1 mentions)
Mef2a, by Santa Cruz Biotechnology (1 mentions)
Odyssey infrared imager, by LI COR (1 mentions)
Odyssey blocking buffer, by LI COR (1 mentions)
2 protocols using sc 313
1
RNA Extraction and Real-Time PCR Analysis
2
Evaluation of Cellular Factors in HTLV
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First, protein levels of MEF-2A and other cellular factors were assessed in Jurkat, MT-2, control and HTLV-infected primary CD4+ T cells by Western blotting. Equal protein quantities from each sample were resolved by SDS-PAGE and transferred to a PVDF membrane. Membranes were blocked for 1 hr at room temperature with Odyssey blocking buffer (Li-Cor Biosciences) and then incubated with antibodies against β-actin (Millipore), p300, p/CAF, and CBP (Abcam), pCREB, and MEF-2A (SC-313, Santa Cruz), HDAC9 (Thermo-Pierce) for 1 hr. Membranes were incubated with IRDye-conjugated secondary antibodies and signals were detected using Odyssey Infrared Imager (Li-Cor). For Co-IP, cells were suspended in ice-cold lysis buffer [Tris–HCl pH 7.4 (25 mM), NaCl (150 mM), NP-40 (1%), EDTA (1 mM), glycerol (5%)] and sonicated. The supernatant was pre-cleared with protein A/G magnetic beads and immunoprecipitation with 2 μg of MEF-2A (Santa Cruz) or Tax (LT-4) antibody followed by Western blotting with same set of antibodies as above.
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