Figure 4.

p97/VCP mediates the HDAC6-dependent HSF1 activation. (A) Human HEK cells stably expressing an anti-HDAC6 shRNA (KD) or the control cell lines (WT) (Kawaguchi et al. 2003; Kovacs et al. 2005) were treated with MG132 (5 μM, up to 18 h), and extracts were prepared after indicated periods of time. The corresponding blots were then probed with the anti-HSF1, anti-HSP70, and anti-tubulin antibodies. (B) HDAC6 knocked-down HEK (KD) and the control (WT) cell lines were treated with control siRNA or siRNA designed against p97/VCP, and cells were treated with 5 μM MG132 during 6 h as indicated. Extracts were prepared and probed with antibodies against HSF1, ubiquitin, p97/VCP, and tubulin.