17-AAG induces cytoplasmic relocalization of FANCA. HeLa/FH-FANCA cells (A-D) and control HeLa cells (E) were treated with 250 nM 17-AAG for the indicated times, and then stained with anti-HA antibody. GM6914 cells stably expressing wild-type FANCA (GM6914/FANCA) were cultured with vehicle (F) or 250 nM 17-AAG (G) for 18 hours, and then stained with anti-FANCA antibody. As a control, GM6914 cells treated with 250 nM 17-AAG for 18 hours were stained with anti-FANCA antibody (H). For optimal visualization of fluorescence signals, exposure time was adjusted. Cell nuclei were visualized with DAPI staining. Images were obtained on an Olympus AX70 microscope equipped with UPlan Apo 20×/0.70 NA and WH10×/22 lenses (Olympus, Tokyo, Japan) using a PXL charged-coupled device camera (model CH1; Photometrics, Osnabruck, Germany).