Optineurin tunes outside-in signaling to regulate lysosome biogenesis and phagocytic clearance in the retina

Blots 1 and 2: Immunoblot of outer segment phagocytosis (rhodopsin) in primary RPE pre-treated or not with 2,4,6-triiodophenol (TIP) (40µM, 30 min or 60 min as indicated) followed by outer segment feeding (~40 OS/cell, 30 min, in the presence of TIP). Loading control: β-tubulin. Lanes 1-3: No treatment, Lanes 4-6: 30 min TIP, Lanes 7-9: 60 min TIP.

Blot 3: NativePAGE of RPE lysates immunoblotted for optineurin. Cells were either untreated (Lanes 1-3) or fed with outer segments (~40 OS/cell, 30 min) (Lanes 4-6), or treated with either mitoChlorodinitrobenzoic acid (MC, 5 µM, 30 min) (Lanes 7-9) or Urolithin A (UA, 100 µM, 24 h) (Lanes 10-12). Arrowhead: ~420 kD optineurin hexamers.