Signaling Pathway Immunoblot Protocol

Antibodies for G6PC (Abcam, Cambridge, MA, USA; ab83690; used at a dilution of 1/500), HNF4α (Abcam, ab181604; used at a dilution of 1/1000), PEPCK (Abcam, ab70358; used at a dilution of 1/1000), and beta-actin (ACTB; Cell Signaling, 4967; used at a dilution of 1/5000) were purchased. Plasmids expressing human H19 (pH19) and empty vector (Vec) were previously described.^{27 (link)} Metformin (ENZO Life Sciences International Inc., Uniondale, NY, USA; ALX-270-432-G005) and DEA (Santa Cruz, Dallas, TX, USA; sc-207632) were purchased. DEA was used at a final concentration of 20 μM.

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Deng J., Mueller M., Geng T., Shen Y., Liu Y., Hou P., Ramillapalli R., Taylor H.S., Paidas M, & Huang Y. (2017). H19 lncRNA alters methylation and expression of Hnf4α in the liver of metformin-exposed fetuses. Cell Death & Disease, 8(12), e3175-.

Publication 2017

ab83690 ab181604 ab70358 Metformin

Antibodies Beta actin Dilution Human Metformin Pepck Plasmids Vector

Corresponding Organization :

Other organizations : Yale University, First Affiliated Hospital of Xi'an Jiaotong University, University Hospital of Bern

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Variable analysis

independent variables

Plasmids expressing human H19 (pH19) and empty vector (Vec)
Metformin (ENZO Life Sciences International Inc., Uniondale, NY, USA; ALX-270-432-G005)
DEA (Santa Cruz, Dallas, TX, USA; sc-207632)

dependent variables

Antibody expression levels of G6PC, HNF4α, PEPCK, and beta-actin (ACTB)

control variables

Antibody dilutions: G6PC (1/500), HNF4α (1/1000), PEPCK (1/1000), and beta-actin (ACTB; 1/5000)

controls

Positive control: Plasmids expressing human H19 (pH19)
Negative control: Empty vector (Vec)

Annotations

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