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. 2012 Sep 28;1(12):1204–1214. doi: 10.1242/bio.20122659

Fig. 2. LX108, R3G3 and R3F2 recognize mitotically phosphorylated GRASP65 in cell lysate by Western blot.

Fig. 2.

(A–D) HeLa cells (lane 1 in each blot), or HeLa cells stably expressing GFP, GFP-tagged rat wild type (WT) GRASP65, or the mG mutant with all 7 phosphorylation sites mutated to alanines (Fig. 1A) (lanes 2–4 in each blot) were analyzed by Western blot using the indicated antibodies. Only S277 (detected by R3G3) is phosphorylated in interphase cells but at a relatively lower level than in mitosis (C). (E) Non-synchronized interphase (Int) or nocodazole-blocked, synchronized mitotic (Mit) HeLa cells expressing WT or mG GRASP65-GFP were analyzed by Western blot, LX108 and R3F2 were able to detect the phospho-protein in WT transfected synchronized cells (lane 2 in each blot).