human free thyroxine ft4 elisa kit  (Cusabio)

Journal: Endocrine-Related Cancer

Article Title: Divergent PTEN–p53 interaction upon DNA damage in a human thyroid organoid model with germline PTEN mutations

doi: 10.1530/ERC-24-0216

Figure Lengend Snippet: Human thyroid organoids resemble human thyroid gland morphology and function. (a) Schematic of thyroid organoid induction protocol. (b) Immunofluorescence (IF) images showing day −1 hiPSCs expressing OCT4 and SOX2, day 0 EBs positive for OCT4 and SOX2, day 9 AFP+ and PAX9+ anterior endoderm cells, day 25 thyroid precursor organoid positive for TTF1 with limited expression of cell death marker-cleaved CAS3 and day 35 mature thyroid organoids that express TPO, PAX8 and TSHR. (c) A panel of IF images capturing TG, TPO, TSHR, and PAX8 expressions in wild-type PTEN thyroid organoids that show a resemblance of subcellular localization (nuclear vs cytoplasm) and follicular morphology when compared to the immunohistochemistry images of normal human thyroid gland taken from the Human Protein Atlas. (d) ELISA-detected thyroid hormone (T4) in the lysates from the thyroid organoid of the three PTEN genotypes (kit sensitivity = 0.6 nmol/L). (e) Western blots of NIS, TTF1, and TSHR in the three genotypes and the human thyroid cell line S-THY 1-3, a positive control. (f) IF panel showing WT/WT , WT/M134R, and WT/G132D mature thyroid organoids positive for TTF1 and TSHR.

Article Snippet: Human Free Thyroxine (fT4) ELISA Kit (MyBioSource, USA; cat. no. MBS266723) was used following the manufacturer’s protocol.

Techniques: Immunofluorescence, Expressing, Marker, Immunohistochemistry, Enzyme-linked Immunosorbent Assay, Western Blot, Positive Control

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