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. 2014 May 22;289(29):19894–19906. doi: 10.1074/jbc.M114.570739

FIGURE 4.

FIGURE 4.

HSF1 isoforms are post-translationally modified. A, Hsf1 isoform ratio in wild type primary fibroblast cell lines. The experiment was performed as described in the legend for Fig. 2. Data are mean (n = 4) ± S.E. HS = heat shock. B, characterization of the established primary cell lines. Wild type (Hsf1+/+) and knock-out (Hsf1−/−) fibroblast lines were analyzed for protein expression levels of HSF2, HSF3, HSF4, HSP70 (Hspa1a/b), HSP40 (Dnajb1), and HSP25 (Hspb1). GAPDH and β-actin (ACTB) were used as loading controls. rec = recovery. C, time course of HSF1 isoform activation. Wild type and knock-out fibroblast lines were transfected with vector (−) and the knock-out line in addition to the four HSF1 isoforms (α, β, γα, γβ). Cells were kept at 37 °C (basal) or heat-shocked and recovered for the indicated time period (0–4 h rec) at 37 °C before Western blot analysis. HSF1 isoforms were detected with anti-HSF1 (Abcam). β-Actin (ACTB) was used as a loading control. ◀ indicates ∼70 kDa.