Skip to main content
. Author manuscript; available in PMC: 2012 Feb 1.
Published in final edited form as: Nat Chem Biol. 2011 Jul 3;7(8):531–537. doi: 10.1038/nchembio.598

Figure 2.

Figure 2

. Characterization of the LID domain. (a) NIH3T3 cells stably expressing YFP-LID were treated with either vehicle or 10 μM MG132 in the presence or absence of 1 μM Shield-1 for 6 h followed by flow cytometry analysis. Lysates from the cells described were resolved by SDS-PAGE and immunoblotted with either anti-YFP or anti-Hsp90 antibodies (Supplementary Fig. 9). (b) Fluorescence microscopy of cells stably expressing the YFP-LID fusion. Cells were treated with either vehicle or 1 μM Shield-1 for 24 h and analyzed using epifluorescence microscopy. HcRed serves as a marker for infection. Insert scalebars represent 10 μm. (c) Cells stably expressing the YFP-LID fusion were treated with various concentrations of Shield-1 (1 μM to 1 pM) and monitored by flow cytometry. (d) The degradation (squares) of YFP-LID was monitored at various times following addition of Shield-1. The recovery (triangles) of YFP-LID was monitored at various time points after depletion of Shield-1 from the culture media. Both experiments were analyzed by flow cytometry. The maximum observed fluorescence intensity for each construct was set to 100%. (e) Cells stably expressing either the YFP-LID or the YFP-19mer fusion proteins were treated with vehicle (−) or with 1 μM Shield-1 for 24 h (+) and analyzed by flow cytometry. The error bars represent the s.d. of the mean based on at least two experiments.