Fig. 2.

Inhibition of HSP90 impairs both HR and NHEJ. (A) HeLa cells stably expressing the DR–GFP reporter were electroporated with either control empty vector or a vector expressing I-SceI and were immediately plated into either DMSO or 250 nM 17-AAG for 24 h. Media was replaced after 24 h (no drug included) and cells were incubated for an additional 24 h before flow-cytometric analysis. Gated cells express GFP, indicating successful HR. Graph depicts number of GFP+ cells in three independent experiments. (B) I-SceI and I-SceI+BcgI digested PCR products. Gray bars in graph represent uncut (0.65 kb) fragment. (C) Summary of total repair capacity and repair pathway distribution in I-SceI transfected cells. Small graphs represent distribution of total repair product (HR+NHEJ). n = 3. Error bars represent SEM; *P < 0.05, **P < 0.01, and ***P < 0.001 (Student’s t test).