(A) HEK293 cell extracts overexpressing LRRK2[Y1699C] and the indicated HA-Rab8A (A) or HA-Rab10 (B) were treated ±150 nM MLi-2 for 90 min and then lysed. The indicated amounts of cell lysates were subjected to either conventional polyacrylamide gel electrophoresis (upper panel) or Phos-tag polyacrylamide gel electrophoresis (lower panel). Proteins on both gels were transferred to nitrocellulose and subjected to identical immunoblot analysis with indicated antibodies (all at 0.5 µg/ml antibody). The open circles on the Phos-tag gel correspond to the position that phosphorylated Rab protein migrates, and the closed circle to the position that the dephosphorylated Rab protein migrates. Immunoblots were developed with ECL.