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. 2017 Nov 30;8:888. doi: 10.3389/fphys.2017.00888

Figure 1.

Figure 1

OR51E2 is expressed in human retinal pigment epithelial cells. (A) The heat map shows the FPKM values for the most abundant ORs (mean FPKM > 0.3) in three whole RPE samples compared to fetal RPE (fRPE), retina supporting tissue (RPE/Choroid/Sclera) and the three neural retina samples (upper panel). The darker colors indicate higher FPKM values and white indicates the absence of any detectable transcripts. Representation of the read coverage of the OR51E2 transcripts detected in primary RPE cells and visualized by the Integrative Genomic Viewer (lower panel). The gray segments indicate the reads that were mapped onto the reference genome. The gene is indicated by the blue bars (exon), and thin line (intron) with an arrowhead that shows the reading direction. Read coverage is shown above (detected and mapped counts/bases at each respective position). (B) Detection of OR51E2 transcripts in primary RPE cells by RT-PCR. Gel electrophoresis of amplicons from primary RPE cell cDNA (+) and no reverse transcriptase cDNA controls (–) to exclude genomic DNA contamination. Premelanosome protein (PMEL) and Retinaldehyde-binding protein 1 (RLBP1) expression identifies RPE cells. The amplification of β-actin (ACTB) served as a control for cDNA quality. (C) Immunofluorescence confocal micrographs of RPE cells labeled with an OR51E2 specific antibody (green) and DAPI to visualize the nuclei (blue). An intracellular staining of OR51E2 can be observed. The bar indicates 50 μm. (D) Plasma membrane localization of OR51E2 in RPE cells verified by surface biotinylation and detection by Western blotting. A representative Western blot of biotinylated membrane of the RPE cells and melanocytes (NHEM), as a positive control, is shown. The cytosolic protein GAPDH served as a control for the enrichment of cell surface proteins and lack of cytosolic proteins. (E) Immunofluorescence confocal micrographs of a histological section of RPE, choroid (Ch) and sclera (S) co-labeled with an OR51E2-specific antibody (red). The bar indicates 50 μm.