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. Author manuscript; available in PMC: 2016 Mar 3.
Published in final edited form as: Structure. 2015 Feb 12;23(3):472–482. doi: 10.1016/j.str.2015.01.003

Figure 5. Posttranslational modifications of Hsc70 residues modulate CHIP-mediated ubiquitination.

Figure 5

(A) Western blot with anti-GST antibody to detect ubiquitination of GST~Hsc70395-646Y611E phosphomimic, GST~Hsc70395-646K561R -dimethyl-mimetic in comparison to wild type (WT) GST~Hsc70395-646 by wild type CHIP1-303.

(B) Pull-down of GST~Hsc70395-646 Y611E and K561R mutant constructs by wild type His6-CHIP1-303 bound to Ni2+ mag-sepharose beads, analyzed by SDS PAGE with Coomassie staining. Lanes indicate the GST~Hsc70395-646 and CHIP1-303 components utilized (INPUT), the final wash fraction (Wash) and the elution fraction (Elute). See also Figure S5.