Luciferase Assays for lncEGFL7OS/EGFL7 Promoter

Luciferase assays were performed as described (Wang et al., 2008a (link)). The putative bidirectional promoter for lncEGFL7OS/EGFL7 was PCR amplified from human DNA and cloned into promoterless PGL3 Basic luciferase vector (Promega). Primers include: plncEGFL7OSup (XhoI): 5’-atcgCTCAGATAGACTCTGATGGCCCAGG-3’ and plncEGFL7OSdn (XhoI): 5’ –atcgCTCAGACCAGCTTGGTGCAGGGAG-3’. 293 T cells in 24-well plates were transfected with 50 ng of reporter plasmids in the presence or absence of increasing amount of Ets1 or Ets1 DNA-binding mutant expression plasmid.

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Zhou Q., Yu B., Anderson C., Huang Z.P., Hanus J., Zhang W., Han Y., Bhattacharjee P.S., Srinivasan S., Zhang K., Wang D.Z, & Wang S. (2019). LncEGFL7OS regulates human angiogenesis by interacting with MAX at the EGFL7/miR-126 locus. eLife, 8, e40470.

Publication 2019

PGL3 Basic luciferase vector

293 t cells Assays Human Luciferase Pgl3 Plasmid Primers Promega Vector

Corresponding Organization :

Other organizations : Tulane University, Boston Children's Hospital, Harvard University, Xavier University, University of Rochester, Oklahoma Medical Research Foundation

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Variable analysis

independent variables

Increasing amount of Ets1 expression plasmid
Increasing amount of Ets1 DNA-binding mutant expression plasmid

dependent variables

Luciferase activity

control variables

293T cells in 24-well plates
50 ng of reporter plasmids

controls

Positive control: Transfection with reporter plasmids in the absence of Ets1 or Ets1 DNA-binding mutant expression plasmid
Negative control: Not explicitly mentioned

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