Journal: Oncotarget

Article Title: Plasma glutamate carboxypeptidase is a negative regulator in liver cancer metastasis

doi: 10.18632/oncotarget.12967

Figure Lengend Snippet: A. Semi-quantitative RT-PCR analysis of DKK4 expression in SK-Hep1 cells treated with PGCP supernatant after PGCP knockdown. ACTB was used as an internal control (top). The signal intensity corresponding to DKK4 was quantified using the ImageJ sofware (bottom). B. Western blot analysis after the DKK4 addition to PGCP knockdown cells. Top, Western blot analysis after transfection with FALG-Mock or FLAG-DKK4, respectively. After 24 h, the culture media including overexpressed DKK4 recombinant proteins was concentrated using an Amicon column. The samples were immunoblotted with anti-DKK4 antibody. Bottom, SK-Hep1 cells were treated with siRNAs (siCont and siPGCP) for 48 h, and secreted DKK4 was added to the culture media. After 24 h, Western blot analysis was performed using the indicated antibodies. ACTB was used as an internal control. C. Detection of phospho-GSK3β (Ser9) and β-catenin after siDKK4 silencing in SK-Hep1 and SNU-387 cells. Western blot analysis was performed using the indicated antibodies. ACTB was used as an internal control. D. Detection of phospho-LRP6 by Western blotting. SK-Hep1 and SNU-387 cells treated with siCont or siPGCP were grown with or without serum. Whole cell lysates were analyzed by Western blotting using the indicated antibodies. ACTB was used as a loading control. E. T4 assay using a Human Free Thyroxine ELISA kit. SK-Hep1 cells treated with siCont or siPGCP were cultured for 48 h. The p value was calculated using Student's t-test (*p<0.05) (top). Quantitative RT-PCR analysis of DKK4 mRNA level after T4 treatment for 24 h. ACTB was used as a loading control. The signal intensity corresponding to DKK4 was quantified using the ImageJ software (bottom). F. Detection of phospho-GSK3β (Ser9) and β-catenin. Western blot analysis was performed after treatment with T4 (final 100 ng/ml) in siRNA-treated SK-Hep1 cells using the indicated antibodies. ACTB was used as an internal control (top). Cell migration assay after T4 treatment. Cells treated with siCont or siPGCP were incubated with T4 (final 100 ng/ml) for 12 h before cell migration assay. DMSO was used as a negative control (bottom).

Article Snippet: The concentration of T4 was measured by the Human Free Thyroxine (FT4) ELISA kit (CSB-E05078h, CUSABIO, Wuhan, China).

Techniques: Quantitative RT-PCR, Expressing, Knockdown, Control, Western Blot, Transfection, Recombinant, Enzyme-linked Immunosorbent Assay, Cell Culture, Software, Cell Migration Assay, Incubation, Negative Control

Journal: PloS one

Article Title: Early postnatal exposure to a low dose of decabromodiphenyl ether affects expression of androgen and thyroid hormone receptor-alpha and its splicing variants in mouse Sertoli cells.

doi: 10.1371/journal.pone.0114487

Figure Lengend Snippet: Figure 1. Serum TH levels in control and decaBDE-exposed mice. Total T3 (A), total T4 (B), FT3 (C) and FT4 (D) levels in the serum of control and decaBDE-exposed mice were measured by ELISA. Each value is the mean ¡ SD of 6 samples per group.

Article Snippet: Similarly, the levels of free T3 (FT3) and free T4 (FT4) in the serum were measured using free tri-iodothyronine indes and free thyroxine ELISA kits (Cusabio Biotech Co., Wuhan, PR China), respectively, according to the manufacturer’s instructions.

Techniques: Control, Enzyme-linked Immunosorbent Assay