Figure 2.
Ligand-induced GPR34 internalization is impaired by mutation. (A) Analysis of subcellular localization of GPR34 and its various mutants by using time-lapse microscopy. Isogenic Flp-InTRex293 cell lines that stably express a single copy of GPR34-GFP or its various mutants were treated with LysoPS (100 µM), and GPR34 expression was monitored over a 30-minute period. Shown is GPR34 expression at selected time points after ligand stimulation. The video record is presented in supplemental Figure 5. (B) The membrane expression in individual cells was quantified at the indicated time by using ImageJ software and normalized to the 0 time point. Comparison between wild-type (WT) GPR34 and its various mutants was performed with Prism 6 nonlinear regression analyses (GraphPad Software), with significant differences indicated. (C) GPR34 immunohistochemistry shows examples of strong (left), moderate (middle), and weak (right) staining.

Ligand-induced GPR34 internalization is impaired by mutation. (A) Analysis of subcellular localization of GPR34 and its various mutants by using time-lapse microscopy. Isogenic Flp-InTRex293 cell lines that stably express a single copy of GPR34-GFP or its various mutants were treated with LysoPS (100 µM), and GPR34 expression was monitored over a 30-minute period. Shown is GPR34 expression at selected time points after ligand stimulation. The video record is presented in supplemental Figure 5. (B) The membrane expression in individual cells was quantified at the indicated time by using ImageJ software and normalized to the 0 time point. Comparison between wild-type (WT) GPR34 and its various mutants was performed with Prism 6 nonlinear regression analyses (GraphPad Software), with significant differences indicated. (C) GPR34 immunohistochemistry shows examples of strong (left), moderate (middle), and weak (right) staining.

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