Journal: Scientific Reports

Article Title: The iodide transporter Slc26a7 impacts thyroid function more strongly than Slc26a4 in mice

doi: 10.1038/s41598-022-15151-4

Figure Lengend Snippet: Serological phenotypes of the deficient mice. ( a ) Serum free thyroxine (FT4), free triiodothyronine (FT3), and thyrotropin (TSH) levels in male wild type (WT), Slc26a7 +/− , and Slc26a7 −/− mice at day 45; n = 4–7 mice of each genotype. One-way analysis of variance (ANOVA) showed significant differences in FT4 and TSH levels (FT4: F (2, 11) = 23.4, p = 0.0001; TSH: F (2, 10) = 22.5, p = 0.0002), but not in FT3 levels ( F (2, 10) = 1.76, p = 0.219). *** p < 0.001 determined by Tukey’s test. ( b ) Serum FT4, FT3, and TSH levels in male WT, Slc26a4 −/− , Slc26a7 −/− , and Slc26a7 del/del Slc26a4 −/− mice at day 90; n = 4–6 mice of each genotype. One-way ANOVA showed significant differences in FT4, FT3, and TSH levels (FT4: F (3, 16) = 22.5, p < 0.0001; FT3: F (3, 14) = 5.29, p = 0.120; TSH: F (3, 14) = 30.3, p < 0.0001). * p < 0.05, *** p < 0.001, **** p < 0.0001 determined by Tukey’s test. FT4 free thyroxine, FT3 free triiodothyronine, TSH thyrotropin, WT wild type.

Article Snippet: Serum free thyroxine (FT4), free triiodothyronine (FT3), and thyrotropin (TSH) concentrations were determined by enzyme-linked immunosorbent assay according to the manufacturer’s instructions (FT4: Mouse Free Thyroxine ELISA Kit CSB-E05080m, CUSABIO, Houston, TX, USA; FT3: Mouse free tri-iodothyronine ELISA kit CSB-E05077m, CUSABIO; TSH: rodent TSH ELISA test kit ERKR7015, Endocrine Technologies, Newark, CA, USA).

Techniques:

Journal: Scientific Reports

Article Title: The iodide transporter Slc26a7 impacts thyroid function more strongly than Slc26a4 in mice

doi: 10.1038/s41598-022-15151-4

Figure Lengend Snippet: Phenotypes of Slc26a7 −/− and Slc26a4 −/− mice on a low iodine diet. ( a ) Urine iodine levels of female Slc26a7 +/− mice with normal and low iodine diets; n = 6 mice in each group. ** p < 0.01 determined by Student’s t test. ( b ) Growth curve of male wild type (WT), Slc26a7 +/− , Slc26a7 −/− , and Slc26a7 −/− mice receiving l -thyroxine (L-T4) replacement from day 21 with a low iodine diet. Note that Slc26a7 −/− mice without thyroid hormone replacement did not survive. ( c ) Serum free thyroxine (FT4) levels in male WT, Slc26a7 +/− , and Slc26a7 −/− mice with normal and low iodine diet at day 21; n = 4–5 mice of each genotype. Two-way analysis of variance (ANOVA) showed significant effects of genotype ( F (2, 20) = 29.92, p < 0.0001). ** p < 0.01, *** p < 0.001 determined by Tukey’s test compared with WT mice with same diet. ( d ) Serum FT4 levels in male WT, Slc26a4 +/− , and Slc26a4 −/− mice that were fed a low iodine diet, at day 90; n = 4–5 mice of each genotype. One-way ANOVA showed no significant differences among the three groups ( F (2, 10) = 0.19, p = 0.83). WT wild type, L-T4 l -thyroxine, FT4 free thyroxine.

Article Snippet: Serum free thyroxine (FT4), free triiodothyronine (FT3), and thyrotropin (TSH) concentrations were determined by enzyme-linked immunosorbent assay according to the manufacturer’s instructions (FT4: Mouse Free Thyroxine ELISA Kit CSB-E05080m, CUSABIO, Houston, TX, USA; FT3: Mouse free tri-iodothyronine ELISA kit CSB-E05077m, CUSABIO; TSH: rodent TSH ELISA test kit ERKR7015, Endocrine Technologies, Newark, CA, USA).

Techniques:

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

Journal: Thyroid

Article Title: A Direct Comparison of Thyroid Hormone Receptor Protein Levels in Mice Provides Unexpected Insights into Thyroid Hormone Action

doi: 10.1089/thy.2019.0763

Figure Lengend Snippet: CRISPR Cas9-mediated knock-in of 2XHA tag. (A) Schematic representation of the mouse Thra and Thrb genes with respective location of HA-coding sequences for ThraHA, Thrb1HA, and Thrb2HA. Open triangles indicate position of insert, solid rectangles and vertical lines represent coding exons, and open rectangles represent untranslated portions of exons. Long introns (horizontal lines) interrupted by double slash. (B) Relative expression of Thra and Thrb in homozygous knock-in mice livers compared with WT C57BL/6. Gene expression levels were determined by qRT-PCR and normalized to β-actin (n > 4). (C) Knock-in animals showed no significant difference in serum thyroid hormone level or TSH compared with WT. Adult male mice (4–6 months old) of each genotype were used for T3 (n = 3), fT4 (n > 6), and TSH (n > 6) measurement. Error bars are SD. One-way ANOVA was used for statistical analysis. ANOVA, analysis of variance; fT4, free thyroxine; HA, hemagglutenin epitope; qRT-PCR, quantitative reverse transcription PCR; SD, standard deviations; tT3, total triiodothyronine; TSH, thyrotropin; WT, wild type.

Article Snippet: Serum total triiodothyronine (tT3) and free thyroxine (fT4) were measured using Mouse Free Thyroxine and Mouse Tri-iodothyronine ELISA Kits (Aviva Systems Biology, San Diego, CA).

Techniques: CRISPR, Knock-In, Expressing, Gene Expression, Quantitative RT-PCR, Reverse Transcription

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