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. 2013 Dec 23;203(6):1043–1061. doi: 10.1083/jcb.201306019

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© 2013 Shahbazi et al.

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Figure 2. — p120 and CLASP2 interact via the N-terminal domain of p120 and the Ser/Arg-rich region of CLASP2. (A) GST-tagged constructs of p120 and CLASP2 used for the in vitro pull-down assays. (B) SDS-PAGE gel showing the purified GST-CLASP2 recombinant proteins stained with Coomassie blue. Asterisks indicate bands of the expected molecular mass. (C) In vitro binding assay of GST-CLASP2 recombinant proteins with purified p120FL and p120ΔN (previously cleaved from GST with the PreScission Protease). (D) In vitro binding of GST-CLASP2 recombinant proteins with purified p120N. (E) Pull-down assay using the recombinant GST-CLASP2 proteins as bait for endogenous p120 present in lysates of mKer treated with calcium for 4 h. (E) GFP-tagged constructs of CLASP2. (F) Pull-down assays using GST-p120N recombinant protein and lysates of 293T cells expressing equal amounts of the GFP-tagged CLASP2 constructs of E. Pulled down proteins were immunoblotted for GFP. (H) Pull-down assay using GST-p120N recombinant protein and lysates of 293T cells expressing equal amounts of either GFP–CLASP2-N1 or GFP–CLASP2-ΔNΔC or GFP–CLASP2-TOG. WB, Western blot.