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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) <t>ELISA-detected</t> 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.
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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) <t>ELISA-detected</t> 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.
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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.

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