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

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Copyright © 2020 Sposini, De Pascali, Richardson, Sayers, Perrais, Yu, Palmer, Nataraja, Reiter and Hanyaloglu

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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LMW compounds induce Gαs activation from FSHR endosomes. (A) TIR-FM images of FLAG-FSHR (red) expressing cells transfected with Nb37-GFP (green) treated with either FSH (10 nM), B3 or T1 (10 μM) for 5 min. Scale bar = 10 μm. (B) Quantification of FSHR endosomes from (A); n = 24 cells/condition in three independent experiments. (C) TIR-FM images (top) of FLAG-FSHR (red), Nb37-GFP (green) and endogenous APPL1 (magenta) in cells stimulated with either FSH, B3 or T1 for 5 min. Scale bar = 3 μm. Line scan analysis of fluorescence intensity (bottom) is shown for one endosome per condition showing FSHR-Nb37-APPL1 colocalization (representative of 233, 302, 316 endosomes for FSH, B1, T3, respectively). (D) Quantification of FSHR-Nb37 endosomes positive for APPL1 after 5-min stimulation with either FSH, B3 or T1; n = 11–12 cells/condition from (C) from three independent experiments.