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. 2019 May 1;317(1):F172–F186. doi: 10.1152/ajprenal.00624.2018

Fig. 2.

Fig. 2.

Surface trafficking of renal olfactory receptors (ORs) visualized by immunohistochemical staining. Live human embryonic kidney-293T cells, transfected with an OR of interest, were stained with rabbit poly-Flag antibody (surface) and then fixed, permeabilized, and stained with mouse mono-Flag antibody (internal). ORs were tagged with the Lucy and Rho tags and coexpressed with receptor-transporting protein 1S (RTP1S) to help promote surface expression. Representative images are shown for each OR. Images in A were taken at a shorter exposure than those in B. ORs that trafficked to the cell surface in >90% of fields of view are indicated by +, whereas +* indicates surface trafficking visible in <50% of fields of view. C: quantification of OR surface expression, plotting the total number of positive cells per field of view. “S” is surface Flag stain (plotted with red circles), “I” is internal Flag stain (plotted with green squares), and “T” is nuclear stain (plotted in blue triangles). Each data point represents a single field of view. Red-to-green ratios are shown with ±SE. Arrowheads indicate cells with positive staining.