SDF-1–induced ERM protein dephosphorylation depends on PLC signaling. (A) Detection of ERM protein phosphorylation by WB in SDF-1–stimulated PBT. Freshly isolated PBTs from a healthy donor were preincubated with PLC inhibitor U73122, its nonfunctional analogue U73433, or PI3-K inhibitor Ly294002 for 10 min and stimulated with/without SDF-1 or PLC activator m-3M3FBS for 45 s. Note that phosphorylated moesin and ezrin are both present in PBT but run as a single band under these conditions. (B) Detection of both moesin and ezrin protein dephosphorylation by WB in SDF-1–stimulated PBT. The samples were prepared as in A but were resolved by running on SDS 4–20% Tris-glycine gels to separate moesin from ezrin. The top panel shows a blot for pERM; the top band is ezrin, and the bottom band is moesin. The middle and bottom panels show WB with antiezrin rabbit polyclonal antibody and antiezrin mouse mAb. (C) Quantitative analysis of data from B using Odyssey software (version 3.0; LI-COR Biosciences). CT, control.