Figure 2 | Scientific Reports

Figure 2

From: Thyrostimulin-TSHR signaling promotes the proliferation of NIH:OVCAR-3 ovarian cancer cells via trans-regulation of the EGFR pathway

Figure 2

Expression of TSHR and thyrostimulin in ovarian cancer cell lines.

(A) The TSHR transcript levels in different ovarian cancer cell lines were compared by real-time PCR quantification. The β-actin level served as a normalized control. (B) TSHR and the various glycoprotein hormone subunit genes, including GPA2 and GPB5 for thyrostimulin and CGA and TSHB for TSH, were PCR-amplified from the cDNA of ovarian cancer cells. The β-actin level served as a loading control. (C) Detection of endogenous thyrostimulin in NIH:OVCAR-3. Concentrated conditioned medium (CM) from NIH:OVCAR-3 cells was subjected to Western blotting using the anti-GPB5 antibody. Conditioned media from NIH:OVCAR-3 cells transfected with FLAG-GPA2/GPB5 and purified thyrostimulin (100 ng) were loaded and served as positive controls. The loading amounts are reflected in Coomassie blue staining. (D) Functional tests of endogenous TSHR in different cell lines. Cells were treated with graded doses of thyrostimulin for 16 h followed by the measurement of the amount of cAMP present in the conditioned media. Data are shown as the mean ± SD.

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