Cells were transfected with 20 nM siRNA targeting RIOK3 (SI02223396; siRIOK3) or AllStars Negative control (1027281; siCon) using HiPerFect reagent (Qiagen, Manchester, UK). Custom siRNA oligonucleotides containing 3 sequences targeting HIF-1α (1 CAAGCAACTGTCATATATA, 2 TGCCACCACTGATGAATTA, 3 TGACTCAGCTATTCACCAA) or HIF-2α (1 TAACGACCTGAAGATTGAA, 2 CAAGCCACTGAGCGCAAAT, 3 TGAATTCTACCATGCGCTA) were purchased from Eurogentec (Southampton, UK). pGIPZ plasmids targeting RIOK3 (Clone ID V2LHS_16875; shRIOK3) or non-silencing control shRNAmir (RHS4743; shCon) were obtained from Thermo Scientific. Lentivirus packaging and titration was carried out as described.38 (link) Cells were transduced at MOI 1 and pools of transduced cells were selected with 1 μg/mL puromycin (Life Technologies) 48 h post-transduction.
Pgipz plasmids
Manufactured by Thermo Fisher Scientific
PGIPZ plasmids are a type of laboratory equipment used for genetic engineering and molecular biology research. They are designed to facilitate the expression and study of target genes in cells. The core function of PGIPZ plasmids is to enable the introduction and controlled expression of specific DNA sequences within a cellular environment, which is a crucial step in various research applications.
Automatically generated - may contain errors
Lab products found in correlation
Hiperfect reagent, by Qiagen (2 mentions)
Puromycin, by Thermo Fisher Scientific (2 mentions)
Si02223396, by Qiagen (2 mentions)
Siriok3, by Qiagen (2 mentions)
Custom sirna oligonucleotides, by Eurogentec (2 mentions)
Clone id v2lhs 16875 shriok3, by Thermo Fisher Scientific (2 mentions)
Rhs4743, by Thermo Fisher Scientific (2 mentions)
Plko 1 plasmids, by Thermo Fisher Scientific (2 mentions)
4 protocols using pgipz plasmids
1
Silencing RIOK3 and HIF Transcription
2
Silencing RIOK3 and HIF Transcription
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Cells were transfected with 20 nM siRNA targeting RIOK3 (SI02223396; siRIOK3) or AllStars Negative control (1027281; siCon) using HiPerFect reagent (Qiagen, Manchester, UK). Custom siRNA oligonucleotides containing 3 sequences targeting HIF-1α (1 CAAGCAACTGTCATATATA, 2 TGCCACCACTGATGAATTA, 3 TGACTCAGCTATTCACCAA) or HIF-2α (1 TAACGACCTGAAGATTGAA, 2 CAAGCCACTGAGCGCAAAT, 3 TGAATTCTACCATGCGCTA) were purchased from Eurogentec (Southampton, UK). pGIPZ plasmids targeting RIOK3 (Clone ID V2LHS_16875; shRIOK3) or non-silencing control shRNAmir (RHS4743; shCon) were obtained from Thermo Scientific. Lentivirus packaging and titration was carried out as described.38 (link) Cells were transduced at MOI 1 and pools of transduced cells were selected with 1 μg/mL puromycin (Life Technologies) 48 h post-transduction.
3
Plasmid Constructs for Brd Knockdown
4
Plasmid Constructs for Brd Knockdown
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pGIPZ plasmids targeting human RNA were purchased from Open Bio-systems. shRNA sequences were subcloned to pH1-IRES-Hygro-GFP reporter vector (a gift from Dr. Ruben Hoya Arias; Ivanova et al., 2006 (link)). For the data shown in Figures 6G–6I , the shBRD4 sequence was subcloned to a pTRIPZ inducible lentiviral vector. pLKO.1 plasmids targeting murine Brd2, Brd3, Brd4, Med1, Med12, and a nontargeting control were purchased from Thermo Scientific. See Supplemental Experimental Procedures for sequences. pLVX-IRES-Oct4tdTomato and pLVX-IRES-tdTomato vector (Figures 3I–3K ) were a kind gift of M. Stadtfeld.
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