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. 2020 Nov 30;11:593492. doi: 10.3389/fphar.2020.593492

FIGURE 4.

FIGURE 4

FSHR requires receptor internalization to induce cAMP signaling in response to either FSH or LMW agonists. (A) Confocal images of FLAG-FSHR stably expressing HEK 293 cells pre-treated with either DMSO or Dyngo-4a (30 μM, 30 min) and stimulated with FSH (10 nM), B3 (10 μM) or T1 (10 μM). Scale bar = 5 μm. (B) Intracellular levels of cAMP measured in cells via HTRF expressing FLAG-FSHR pre-treated with either DMSO or Dyngo-4a (30 μM, 30 min) and stimulation either DMSO, FSH (10 nM, 5 min), B3 or T1 (10 μM, 5 min). Data are expressed as cAMP levels normalized to FSH treatment (DMSO). Two-way ANOVA: **p < 0.01, ****p < 0.0001. (C) Percentage decrease in cAMP levels following Dyngo-4a pre-treatment calculated from (B). n = 3 independent experiments.