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. 2016 Nov;186(11):2857–2868. doi: 10.1016/j.ajpath.2016.06.022

Figure 3.

Figure 3

Nuclear accumulation and phosphorylation of HSF1 in FA-treated human cells. A and B: H460 (A) and IMR90 (B) cells were treated (FA and MG132 for 3 hours, 43°C for 1 hour) and immunostained for HSF1. C: Western blots for HSF1 and Ser326-phosphorylated HSF1 in protein extracts prepared from H460 cells using different lysis buffers. FA treatments were for 3 hours. Tubulin was used as a loading control. D: HSF1 phosphorylation and K48-Ub of soluble cellular proteins (1% NP40 extraction) after H460 treatments with FA and MG132. E: Representative images of IMR90 normal human cells immunostained for Ser326–p-HSF1. Cells were exposed to FA or MG132 for 3 hours, and 43°C for 1 hour. FA, formaldehyde; HSF1, heat shock factor 1; K48-Ub, K48-polyubiquitination; p-HSF1, phosphorylated HSF1.