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. 2018 Apr 2;217(4):1269–1285. doi: 10.1083/jcb.201708116

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© 2018 Grousl et al.

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Figure 4. — Hsp42 PrLD is essential for mCherry–VHL interaction. Coimmunoprecipitation of Hsp42 WT and deletion mutants (all FLAG-tagged) and mCherry-myc-VHL. Cells were grown at control conditions (30°C) or stressed (37°C for 45 min + MG132) before extract preparation. Hsp42 proteins were precipitated using anti-FLAG agarose, and the amount of bound Hsp42 and mCherry-myc-VHL was quantified by immunoblot analysis using anti-FLAG and anti-myc antibodies. Fractions were analyzed for the content of FLAG-tagged proteins and mCherry–VHL interaction (anti-myc). The sizes of Hsp42 proteins are indicated (arrows). Asterisks indicate coeluting mouse IgG heavy and light chains. VHL input controls are provided. The dashed line indicates intervening lanes have been spliced out. IP, immunoprecipitation.