Western blot analysis of cellular signaling

Western blot assay was performed using whole cell lysates from either liver tissue or hepatocytes, or media as previously described (4 (link)). Membranes were incubated overnight using the following antibodies: HMGB1 (Abcam) and β-actin (Sigma); phospho-p38, p38, phospho-JNK, JNK, ERK, phospho-ERK, p65, phospho-p65, PARP-1, acetyl-histone H3, acetyl-histone H4, and phospho-histone H2A.X (Cell Signaling); PAR (BD bioscience).

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Huang H., Nace G.W., McDonald K.A., Tai S., Klune J.R., Rosborough B.R., Ding Q., Loughran P., Zhu X., Beer-Stolz D., Chang E.B., Billiar T, & Tsung A. (2014). Hepatocyte specific HMGB1 deletion worsens the injury in liver ischemia/reperfusion: A role for intracellular HMGB1 in cellular protection. Hepatology (Baltimore, Md.), 59(5), 1984-1997.

Publication 2014

HMGB1 β-actin phospho-JNK phospho-ERK phospho-p65 PARP-1 acetyl-histone H3 acetyl-histone H4 phospho-histone H2A.X

Actin Antibodies Hepatocytes Histone h2a x Histone h3 Histone h4 Hmgb1 Membranes Parp 1 Tissue Western blot assay

Corresponding Organization : University of Pittsburgh Medical Center

Other organizations : University of Chicago

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Variable analysis

independent variables

Tissue/cell type (liver tissue or hepatocytes)

dependent variables

Protein expression levels of HMGB1, β-actin, phospho-p38, p38, phospho-JNK, JNK, ERK, phospho-ERK, p65, phospho-p65, PARP-1, acetyl-histone H3, acetyl-histone H4, and phospho-histone H2A.X
Presence of PAR

control variables

Western blot assay protocol as previously described (4)

controls

No positive or negative controls were explicitly mentioned.

Annotations

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