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Figure 1. Schematic representation of TSHR

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1 Figure 1. Schematic representation of TSHR
Figure 1. Schematic representation of TSHR. The five mutations reported in this series of patients and their locations are illustrated. C41S and P162A are located in receptor ectodomain, whereas T655Δ causes the formation of a nonsense codon at 656 and the loss of TM7 and the intracellular tail of the receptor. The mutations L467P and C600R are novel, and they are located in TM2 and at the intracellular end of TM5, respectively. From: Germline Mutations of TSH Receptor Gene as Cause of Nonautoimmune Subclinical Hypothyroidism J Clin Endocrinol Metab. 2002;87(6): doi: /jcem J Clin Endocrinol Metab | Copyright © 2002 by The Endocrine Society

2 Figure 2. Flow cytometry analysis
Figure 2. Flow cytometry analysis. A, Analysis with BA8 monoclonal antibody (mAb), directed against a conformational epitope in the extracellular domain of TSHR (23 ), of intact or permeabilized COS7 cells transfected with either empty pTarget vector or WT-, L467P-, C600R-, or T655Δ-TSHR. B, Analysis with 3G4 mAb, directed against a linear epitope in the extracellular domain of TSHR (23 ), of intact or permeabilized COS7 cells transfected with either empty pTarget vector or WT-, C41S-, L467P-, or C600R-TSHR. From: Germline Mutations of TSH Receptor Gene as Cause of Nonautoimmune Subclinical Hypothyroidism J Clin Endocrinol Metab. 2002;87(6): doi: /jcem J Clin Endocrinol Metab | Copyright © 2002 by The Endocrine Society

3 Figure 3. Basal cAMP accumulation in COS7 or HEK293T cells transiently transfected with WT- (n = 7 or 4, respectively, for COS7 or HEK293T), C41S- (n = 1 in both cell types), L467P- (n = 4 in both), C600R- (n = 4 in both), or T655Δ-TSHR (n = 4 or 3). Each experiment was performed in triplicate wells. Results (mean ± se) are expressed as percentage above basal cAMP accumulation in mock transfected cells. From: Germline Mutations of TSH Receptor Gene as Cause of Nonautoimmune Subclinical Hypothyroidism J Clin Endocrinol Metab. 2002;87(6): doi: /jcem J Clin Endocrinol Metab | Copyright © 2002 by The Endocrine Society

4 Figure 4. Representative hormone-induced cAMP accumulation in COS7 cells transiently transfected with WT-TSHR (•) or empty vector (○) in presence of increasing concentrations of bTSH. From: Germline Mutations of TSH Receptor Gene as Cause of Nonautoimmune Subclinical Hypothyroidism J Clin Endocrinol Metab. 2002;87(6): doi: /jcem J Clin Endocrinol Metab | Copyright © 2002 by The Endocrine Society

5 Figure 5. Stimulation with 10 U/liter bTSH of cAMP accumulation in COS7 or HEK293T cells transiently transfected with empty pTarget vector (n = 7 or 3, respectively, in COS7 or HEK293T), or WT- (n = 7 or 3), C41S- (n = 2 in both cell types), L467P- (n = 7 or 3), C600R (n = 7 or 3), and T655Δ-TSHR constructs (n = 5 or 3). Each experiment was performed in triplicate wells. Results (mean ± se) are expressed as percentage above nonstimulated cells. From: Germline Mutations of TSH Receptor Gene as Cause of Nonautoimmune Subclinical Hypothyroidism J Clin Endocrinol Metab. 2002;87(6): doi: /jcem J Clin Endocrinol Metab | Copyright © 2002 by The Endocrine Society

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