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Methods Main Schwann mobile civilizations Computer mouse button SCs (mSCs) ended up ready utilizing a changed Brockes strategy [28]. Tissues were pure and cultured since defined just before [29]. Briefly, sciatic nerves were dissected from neonatal (postnatal morning Several (P3)) C57BL/6 #links# these animals as well as ingested together with 3.05% collagenase (Worthington, Lakewood, NJ, USA) alo
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Plemental Fig. S2). In contrast, I54A 32 was degraded slightly faster than wild type at 37 and even faster at 42 (Fig. 1 and supplemental Fig. S2).JOURNAL OF BIOLOGICAL CHEMISTRYChaperone Binding toFIGURE 1. In vitro degradation of 32 by ClpYQ and FtsH proteases. 32 (2.4 g) was incubated with ClpQ (1.0 g) and ClpY (2.5 g), or 32 (1.0 g) was incubated with FtsH (2.0 g), in a reaction mixture (6
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Precipitated by adding protein A-Sepharose. After subjecting the precipitate to SDS-PAGE, 32 and DnaK were detected by CBB staining, and DnaJ was detected by immunoblotting with anti-DnaJ serum. Pull-down assays for the interaction between 32 and DnaKHis or DnaJ-His were performed using Ni2 -NTA-agarose. DnaK-His was preincubated in Buffer D (20 mM Tris-HCl (pH 7.5), 100 mM KCl, 10 glycerol, 5 mM
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