ChIP-PCR Assay for EGR1-PFKFB3 Interaction

The ChIP-PCR procedure was similar to previously described procedures [30 (link)]. ChIP DNA from anti-EGR1 (ab55160, Abcam)-treated cells was used to examine the association between EGR1 and PFKFB3. DNA from anti-IgG antibody-treated cells served as the control. Purified DNA was used to analyze the PFKFB3 proximal promoter region by real-time PCR. The PFKFB3 primers were as follows: forward: 5′-TGTGAAAACCAGATGCCAGC-3′; and reverse: 5′-GGACTTGAACTGAGCCTTGC-3′. The relative amplification of the promoter sequence of the gene was calculated using the 2^−ΔΔCT method.

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Song C., Wang S., Fu Z., Chi K., Geng X., Liu C., Cai G., Chen X., Wu D, & Hong Q. (2022). IGFBP5 promotes diabetic kidney disease progression by enhancing PFKFB3-mediated endothelial glycolysis. Cell Death & Disease, 13(4), 340.

Publication 2022

ab55160

Anti dna antibody Cells Chip Egr1 Gene amplification Primers Real time pcr

Corresponding Organization :

Other organizations : Chinese People's Liberation Army, Chinese PLA General Hospital

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Variable analysis

independent variables

Treatment with anti-EGR1 antibody (ab55160, Abcam)

dependent variables

Association between EGR1 and PFKFB3 as measured by ChIP-PCR

control variables

Treatment with anti-IgG antibody

controls

Treatment with anti-IgG antibody

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