Fig. 6.
Determination of Gq/11 signaling via NFAT reporter gene assay of TSHR and MCT8 co-expressed in different ratios. HEK293 cells were co-transfected with TSHR and MCT8, for control, TSHR was co-transfected with a non-interacting control (MC4R, [31]) in the indicated ratios with a starting point of equimolar amounts of both plasmids as it is used for Figs. 1 and 4. Two days after transfection, IP3 formation (NFAT-Luc, via reporter gene assay) was analyzed in the absence or presence of 100 mU/mL bTSH, stimulation of TSHR:MCT8 is indicated in red bars. Data of four independent experiments performed in triplicates are shown. Comparison was performed between TSHR + control versus TSHR + MCT8. Values represent mean + SEM. *p ≤ 0.05, ***p ≤ 0.001, ****p ≤ 0.0001 (unpaired t test with Welch’s correction)