Fig 1: Representative images of the DICER1 immunohistochemistry staining and summary of the staining results. (A and B) The staining of DICER1 in two cases of FTC shows positive staining of the corresponding normal tissue and negative staining in the central parts of the tumour. The tumour tissue, in general, showed weaker DICER1 immunoreactivity compared to the corresponding normal thyroid tissue. Magnification ×400. (C) Summarized results from the immunohistochemical scoring of normal thyroid tissue, follicular thyroid carcinomas (FTC) and Hürthle cell carcinomas (HuCC). A full colour version of this figure is available at https://doi.org/10.1530/ERC-19-0446.
Fig 2: Kaplan–Meier survival curves in follicular thyroid carcinoma (FTC) and Hürthle cell carcinoma (HuCC) patients. (A) The overall survival in low vs high DICER1 expression. (B) The disease-free survival in low vs high DICER1 expression. (C) The disease-specific survival (DSS) in low vs high DICER1 expression. (D) DSS in FTC vs HuCC. P values were calculated using the Log-rank test. Significant P values are in bold. A full colour version of this figure is available at https://doi.org/10.1530/ERC-19-0446.
Fig 3: GABPA knock down in FTC-238 cell line. (A) GABPA mRNA expression in the knock down cells (siGABPA#1 and siGABPA#2) is significantly reduced compared to control cells. (B) DICER1 mRNA expression in the knock down cells (siGABPA#1 and siGABPA#2) is significantly reduced compared to control cells. (C) Confirmation of reduced levels of GABPA and DICER1 after GABPA knock down. Estimated protein sizes are indicated to the right in kilodaltons (kDa). P values were calculated with Student’s t-test. Significant P values are in bold.
Fig 4: Chromatin immunoprecipitation assay of the binding of GABPA to the DICER1 promoter. (A) Gel electrophoresis showing DNA bands from DICER1 promoter PCR product. Lane 1 shows band from 2% input DNA, lane 2 shows band from pulled down DNA using GABPA antibody and lane 3 shows PCR product from cross-linked chromatin DNA incubated with normal rabbit IgG. (B) Chromatogram from the GABPA antibody pulled down DNA and subsequent amplification and sequencing of the DICER1 promoter using the reverse primer as sequencing primer. The sequence was blasted with a 100% match in the DICER1 promoter. A full colour version of this figure is available at https://doi.org/10.1530/ERC-19-0446.
Fig 5: The age dependent differences in DICER1 expression and survival. (A) DICER1 mRNA expression in patients >25 years of age vs <25 years of age. (B) The disease-free survival in patients >25 years of age vs <25 years of age. (C) Correlation of DICER1 and GABPA mRNA expression in patients <25 years of age. (D) Correlation of DICER1 and GABPA mRNA expression in patients >25 years of age. A full colour version of this figure is available at https://doi.org/10.1530/ERC-19-0446.
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