Immunohistochemical and Immunofluorescent Analyses

IHC was performed on formalin-fixed paraffin-embedded (FFPE) sections as described (20 (link)) using anti-AEG-1 (chicken, in-house) and anti-PCNA (Cell signaling #13110) antibodies. IF was performed on FFPE sections using anti-AEG-1 (chicken, in-house) and F4/80 (AbD Serotec #MCA497RT) antibodies. Hepatocytes were cultured in collagen-1 coated 4-chamber slides and IF was performed using antibody against γ-H2AX (Cell signaling #5438). LSEC were cultured in 8-chamber slides and IF was performed using anti-Stabilin-2 antibody (MBL International #D317–3). For IHC, images were analyzed using an Olympus microscope. For IF images were analyzed using a Zeiss confocal laser scanning microscope.

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Robertson C.L., Mendoza R.G., Jariwala N., Dozmorov M., Mukhopadhyay N.D., Subler M.A., Windle J.J., Lai Z., Fisher P.B., Ghosh S, & Sarkar D. (2018). Astrocyte elevated gene-1 (AEG-1) regulates macrophage activation in hepatocellular carcinogenesis. Cancer research, 78(22), 6436-6446.

Publication 2018

#MCA497RT confocal laser scanning microscope

Anti antibody Antibodies Antibody Chicken Collagen 1 Confocal laser scanning microscope Embedded paraffin Formalin Hepatocytes Microscope Pcna

Corresponding Organization : Virginia Commonwealth University Medical Center

Other organizations : The University of Texas Health Science Center at San Antonio

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Variable analysis

independent variables

Antibody used for IHC (anti-AEG-1 and anti-PCNA)
Antibody used for IF (anti-AEG-1, F4/80, γ-H2AX, anti-Stabilin-2)

dependent variables

Protein expression levels of AEG-1 and PCNA (measured by IHC)
Localization of AEG-1, F4/80, γ-H2AX, and Stabilin-2 (measured by IF)

control variables

Formalin-fixed paraffin-embedded (FFPE) tissue sections
Hepatocytes cultured in collagen-1 coated 4-chamber slides
LSEC cultured in 8-chamber slides

controls

Positive control: None specified
Negative control: None specified

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