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. 2000 Sep 12;97(20):10757–10762. doi: 10.1073/pnas.190062797

Figure 3.

Figure 3

Transactivation by Gal4-LZIP requires functional HCF-1. (A) Temperature-dependent transactivation by Gal4-LZIPN154. Subconfluent tsBN67 cells were cotransfected with expression plasmids (250 ng) encoding Gal4-LZIPN154, Gal4-Sp1, and Gal4-c-jun together with the p5xGal-E1B-luc reporter gene (500 ng) and incubated at 33.5°C or 39.5°C for 40 h. Because Gal4-Sp1 and Gal4-c-jun are much weaker activators than Gal4-LZIPN154, activity at 39.5°C is plotted relative to the activity of the same activator at 33.5°C. (B) Transactivation by Gal4-LZIPN154 is restored in tsBN67 cells stably expressing wild-type HCF-1 (tsBN67-R). (C) Time course analysis of tsBN67 cells. Transfected cells were maintained at 33.5°C for 2 h after electroporation with p5xGal-E1B-luc (500 ng) and Gal4-LZIPN154 (250 ng), and then half of the culture was shifted to 39.5°C (time = 0). Aliquots of cells were then collected at 4-h intervals and assayed for luciferase activity. Values represent the means of three assays. (D) Activation by wild-type and HBMKO versions of Gal4-LZIPN92 assayed in tsBN67 cells maintained at 33.5°C.