Figure 5. Crosslinking CRFB1 keeps receptor molecules clustered despite caveolin depletion.

ZFL cells were co-transfected with MO and expression plasmid via nucleofection and allowed to recover/adhere to cell culture plates for ∼6 hr prior to addition of crosslinking reagent. The crosslinking reaction was performed according to the manufacturer’s procedures. Cells were subsequently replenished with media and returned to the incubator for 24 hr post crosslinking. Scale bars, 1 µm. A) Cells transfected with Cav-1b MO/CRFB1 without crosslinking treatment show dispersed receptor molecules. B) Cells transfected with Control MO/CRFB1 with crosslinking clearly show clustered receptor molecules. C) Cells transfected with Cav-1b MO/CRFB1 with crosslinking. This demonstrates that despite depletion of Cav-1b, receptor molecules remain clustered. D) Pair correlation analysis confirms that with crosslinking, CRFB1 remains clustered despite Cav-1b depletion. Values of g(r) in cells with Cav-1b KD are similar to that for Controls (n≥8 cells per treatment).