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Pwzl blast myc

Manufactured by Addgene
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The PWZL-Blast-myc is a laboratory equipment product. It is designed for cell culture and molecular biology applications. The core function of this product is to enable the expression and detection of the myc protein tag in cells. No further details on the intended use or application of this product are provided.

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Lab products found in correlation

Pbabe puro ras v12, by Addgene (2 mentions) Control sirna, by Merck Group (1 mentions) Lipofectamine ltx plus reagent, by Thermo Fisher Scientific (1 mentions) P3xflag myc cmv 26, by Merck Group (1 mentions) Lenticrispr v2, by Addgene (1 mentions) Pbabe yap, by Addgene (1 mentions) Fusion sl4, by Vilber (1 mentions)

6 protocols using pwzl blast myc

1

Modulation of Prostate Cancer Regulators

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Human SREBP-2 cDNA was subcloned into p3XFLAG-myc-CMV-26 (Sigma-Aldrich) at NotI/XbaI restriction enzyme sites to generate p3XFLAG-SREBP-2 expression construct. LNCaP and LAPC4 cells were transfected with either p3XFLAG-SREBP-2 or empty vector as a control using Lipofectamine LTX Plus reagent (Life Technologies). For the SREBP-2 shRNA-mediated knockdown study, non-targeting control (pLKO.1, empty vector) or SREBP-2 shRNA lentiviral particles (Sigma-Aldrich) were used to infect CWR22Rv1 or C4-2B cells. In order to generate c-Myc-overexpressing PCa cell clones, retroviruses that carried either pWZL-Blast-Myc (Addgene) or control vector (pWZL-Blast) were used. For the silencing of c-Myc, cells were transfected with c-Myc siRNA-1, c-Myc siRNA-2 or control siRNA (Sigma-Aldrich) using DharmaFECT 2 transfection reagent (Fisher Scientific Dharmacon).
Li X., Wu J.B., Li Q., Shigemura K., Chung L.W, & Huang W.C. (2016). SREBP-2 promotes stem cell-like properties and metastasis by transcriptional activation of c-Myc in prostate cancer. Oncotarget, 7(11), 12869-12884.
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2

Generating Stable Cell Lines with Lenti- and Retroviral Constructs

Cited 2 times
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Lenti- or retro-viral plasmids pLKO.1 shSox2 HM a (#26353), pLKO.1 shSox2 3H b (#26352), pWZL-Blast-MYC (#10674), and pBABE-YAP (#15682) were purchased from Addgene, Cambridge, MA (Bass et al., 2009 (link); Boehm et al., 2005 (link); Overholtzer et al., 2006 (link)). Mouse Sox2 sgRNA sequences were inserted into LentiCrispr v2 (#52961, Addgene) (Ahnfelt-Ronne et al., 2012 (link)). Specific sgRNA sequences were listed in STAR Methods. Lentiviral and retroviral productions were performed as previously described (Zhang et al., 2014 (link)). To generate stable MYC or YAP overexpressing lines, primary PDAC cells were first selected with blasticidin or puromycin after infection with viruses carrying these genes.
Murakami S., Nemazanyy I., White S.M., Chen H., Nguyen C.D., Graham G.T., Saur D., Pende M, & Yi C. (2019). A Yap-Myc-Sox2-p53 Regulatory Network Dictates Metabolic Homeostasis and Differentiation in Kras-driven Pancreatic Ductal Adenocarcinomas. Developmental cell, 51(1), 113-128.e9.
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3

Lentiviral and Retroviral Vector Production

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Lentiviral vectors were packed with psPAX2 and pMD2.G in 293T cells with polyethylenimine (PEI) transfection reagent. Retroviral vectors pBabe-puro Ras V12 (Addgene #1768) and pWZL-Blast-myc (Addgene #10674) were packed with pEco (Clontech) in 293T cells. The viral solutions were 30-fold concentrated by PEG6000 precipitation, then aliquot and stored at −80°C.
Li W., Hu J., Shi B., Palomba F., Digman M.A., Gratton E, & Jiang H. (2020). Biophysical properties of AKAP95 protein condensates regulate splicing and tumorigenesis. Nature cell biology, 22(8), 960-972.
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4

Lentiviral and Retroviral Vector Production

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Lentiviral vectors were packed with psPAX2 and pMD2.G in 293T cells with polyethylenimine (PEI) transfection reagent. Retroviral vectors pBabe-puro Ras V12 (Addgene #1768) and pWZL-Blast-myc (Addgene #10674) were packed with pEco (Clontech) in 293T cells. The viral solutions were 30-fold concentrated by PEG6000 precipitation, then aliquot and stored at −80°C.
Li W., Hu J., Shi B., Palomba F., Digman M.A., Gratton E, & Jiang H. (2020). Biophysical properties of AKAP95 protein condensates regulate splicing and tumorigenesis. Nature cell biology, 22(8), 960-972.
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5

Modeling Oncogenic Transformation in MEFs

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Mouse embryonic fibroblasts were plated in 6-well plates at a density of 250'000 cells per well and transduced with lentiviral particles carrying shRNA constructs for Luciferase or Xpg (shXpg-A). Cells were then irradiated (8 Gy), followed by co-transfection with plasmids carrying H-Ras (pWZL Blast H-Ras G12V E37G, Addgene #12277) and cMyc (pWZL Blast myc, Addgene #10674), respectively. In a control well, transfected cells were cultured in presence of Blasticidin to control for transfection efficiency. After 15 days, cells were washed twice with 1xPBS, fixed with ice-cold methanol and stained with 0.5% crystal violet. Plates were photographed in a Fusion-SL4 (Vilber).
Avila A.I., Illing A., Becker F., Maerz L.D., Morita Y., Philipp M, & Burkhalter M.D. (2016). Xpg limits the expansion of haematopoietic stem and progenitor cells after ionising radiation. Nucleic Acids Research, 44(13), 6252-6261.
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6

NELFE Overexpression in HHT4 Cells

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pLKO.1 shNELFE and pLKO.1 shCtrl were purchased from OpenBiosystems (TRC lentivirus). Lenti-C-NELFE mGFP tagged ORF or vector were purchased from Origene. pBABE-zeo-largeTgenomic (Addgene #1778) and pWzl-Blast-Myc (Addgene #10674) were transfected into HHT4 cells followed by selection with puromycin and blasticidin, respectively to generate stable cell lines. HHT4-SV40 cells were transfected with NELFE or vector lentivirus at MOI 5 (NELFE mGFP tagged ORF, Origene). See also Key Resources Table for list of plasmids used.
Dang H., Takai A., Forgues M., Pomyen Y., Mou H., Xue W., Ray D., Ha K.C., Morris Q.D., Hughes T.R, & Wang X.W. (2017). Oncogenic activation of the RNA binding protein NELFE and MYC signaling in hepatocellular carcinoma. Cancer cell, 32(1), 101-114.e8.
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