Figure 5.

Transient receptor potential vanilloid channel, member 6 (TRPV6) localization to the plasma membrane increased after treatment with zymosan, a reagent that is phagocytized by the retinal pigment epithelium. Donor R cells were plated (passage 151) onto Permanox four-well chamber slides and grown for 7 days under standard culture conditions. Monolayers were rinsed free of media with standard buffer and equilibrated in that buffer for 30 min at 37 °C. Monolayers were exposed to either standard buffer (Control, upper panel) or standard buffer plus 2 mg/ml zymosan (Zymosan, lower panel) for a further 10 min at 37 °C. Reaction was stopped by formaldehyde fixation, and immunocytochemistry was performed. Primary and secondary antibodies were those used in Figure 4, bottom panel (TRPV6). Nuclei were stained with TO-PRO-3. The scale bar in panel B represents 10 μm and applies to both panels.