![(A) HEK293 cell extracts overexpressing LRRK2[Y1699C] and the indicated HA-tagged Rab protein were treated ±150 nM MLi-2 for 90 min and then lysed. The cell lysates (0.1 µg) were immunoblotted with the indicated MJFF-pRab10 hybridoma clones (all at 0.5 µg/ml antibody). (B) The indicated wild-type and knock-out A549 cells were treated ±100 nM MLi-2 for 90 min and then lysed. The cell lysates (20 µg) were immunoblotted with the indicated MJFF-pRab10 hybridoma clones as well as shown total antibodies (all at 0.5 µg/ml antibody). (C) The indicated amounts of recombinant LRRK2-phosphorylated Rab10 were subjected to immunoblotting with the specified MJFF-pRab10 hybridoma clones as well as the total RAB10 antibody purchased from Cell Signaling Technology (CST) (all at 0.5 µg/ml antibody). (D) LRRK2[R1441G] MEFSs were treated ±100 nM MLi-2 for 90 min and then lysed. The cell lysates (5 µg) were immunoblotted with the indicated MJFF-pRab10 hybridoma clones (all at 0.5 µg/ml antibody). (E) The indicated littermate wild-type and knock-in or knock-out MEF cells were treated ±100 nM MLi-2 for 90 min and lysed. The cell lysates (5 µg) were immunoblotted with the indicated antibodies and the membranes developed using the Odyssey CLx scan Western Blot imaging system (all at 1 µg/ml antibody). Quantitation of immunoblots was undertaken by analysing pRab10/tubulin (Ei). Immunoblots were developed with LI-COR.](https://port.silverchair-cdn.com/port/content_public/journal/biochemj/475/1/10.1042_bcj20170802/2/bcj-2017-0802_04.jpeg?Expires=1716885382&Signature=S45KfjH33-FnODTnEZDSrMgsDk~wgXY9WhoqVgiTis4ZUdVyPTdgncoKZ186HbV3UX~ucvePrwgHu-tCjwAJJqYl~mT1j3ylIHG-XhlZSxrwBk00lxN5zRSDnudK7H0CIaE7F1~mgjkFU-QtoJwCxATQ8ub8k-ubyieIwePyP-xZyrPReoOD2blKzu2fEWP63Xn3hiI5ysq4iaZG04KEuH0~~seH7jqMy9DlsPnRfnHcxS7saNKNx3Jo0q-BAZtTb7izWUqyckDf6gszru2qITp39rToXn6QJQEZfGqBf3QR5JgZILO7g-7b84KSK-MdixI0VEBDjwccjfd5mu3xjw__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
(A) HEK293 cell extracts overexpressing LRRK2[Y1699C] and the indicated HA-tagged Rab protein were treated ±150 nM MLi-2 for 90 min and then lysed. The cell lysates (0.1 µg) were immunoblotted with the indicated MJFF-pRab10 hybridoma clones (all at 0.5 µg/ml antibody). (B) The indicated wild-type and knock-out A549 cells were treated ±100 nM MLi-2 for 90 min and then lysed. The cell lysates (20 µg) were immunoblotted with the indicated MJFF-pRab10 hybridoma clones as well as shown total antibodies (all at 0.5 µg/ml antibody). (C) The indicated amounts of recombinant LRRK2-phosphorylated Rab10 were subjected to immunoblotting with the specified MJFF-pRab10 hybridoma clones as well as the total RAB10 antibody purchased from Cell Signaling Technology (CST) (all at 0.5 µg/ml antibody). (D) LRRK2[R1441G] MEFSs were treated ±100 nM MLi-2 for 90 min and then lysed. The cell lysates (5 µg) were immunoblotted with the indicated MJFF-pRab10 hybridoma clones (all at 0.5 µg/ml antibody). (E) The indicated littermate wild-type and knock-in or knock-out MEF cells were treated ±100 nM MLi-2 for 90 min and lysed. The cell lysates (5 µg) were immunoblotted with the indicated antibodies and the membranes developed using the Odyssey CLx scan Western Blot imaging system (all at 1 µg/ml antibody). Quantitation of immunoblots was undertaken by analysing pRab10/tubulin (Ei). Immunoblots were developed with LI-COR.
