Comprehensive Liver Cell Culture Protocol

Cell culture medium (DMEM/F12), OPTI-MEM, Lipofectamine 2000 and TRIzol reagent were from Life Technologies (San Diego, CA, USA). Antibody against β-actin was obtained from Santa Cruz Biotech (Santa Cruz, CA, USA). Mismatch RNA (mmRNA) was described previously [16 (link)]. Brf1 antibody was from Bethyl laboratories Inc (Montgomery, TX, USA). The sequences of primers and Brf1 siRNA were described in (Supplementary Table S3) [20 (link)]. Tumor stem cells of mouse liver (TSCML) were kindly provided by Dr. K. Machida at USC (University of Southern California). AML-12 cell line was from ATCC. HepG2-ADH cell line was kindly provided by Dr. D.L. Clemens (University of Nebraska). The animal experiments were conducted in accordance with the approved institutional animal care and use committee protocol at USC.

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Zhong Q., Xi S., Liang J., Shi G., Huang Y., Zhang Y., Levy D, & Zhong S. (2015). The significance of Brf1 overexpression in human hepatocellular carcinoma. Oncotarget, 7(5), 6243-6254.

Publication 2015

Lipofectamine 2000 TRIzol reagent Antibody against β-actin Brf1 antibody AML-12 cell line

Actin Antibody Cell Cell culture Cell line Culture medium Hepg2 cell Institutional animal care and use committee Lipofectamine 2000 Mouse Primers Sirna Stem cells Trizol Tumor of liver

Corresponding Organization : Sun Yat-sen University Cancer Center

Other organizations : Shantou University, Shantou University Medical College, University of Southern California

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Variable analysis

independent variables

Mismatch RNA (mmRNA)
Brf1 antibody
Brf1 siRNA

dependent variables

β-actin expression

control variables

Cell culture medium (DMEM/F12)
OPTI-MEM
Lipofectamine 2000
TRIzol reagent
Tumor stem cells of mouse liver (TSCML)
AML-12 cell line
HepG2-ADH cell line

controls

Positive control: Antibody against β-actin
Negative control: Not explicitly mentioned

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