(A) CHO cells transiently transfected with mCherry-D4D434A and D4D434A, A463W and the plasma membrane marker, Pleckstrin homology domain of phospholipase C δ (PH-PLC δ) were examined using spinning-disc confocal microscopy. (B) Live-cell images were acquired of cells expressing the same probes as in (A) following incubation with 10 mM methyl-β-cyclodextrin (mβCD) for 20 min to extract plasmalemmal cholesterol. Scale bar, 10 µm. (C) Quantitation of the plasmalemmal enrichment of the mCherry signal seen in (A) and (B). means ± std. dev. n = 20.