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. Author manuscript; available in PMC: 2015 Mar 15.
Published in final edited form as: J Immunol. 2014 Feb 12;192(6):2892–2903. doi: 10.4049/jimmunol.1302847

Figure 4. Foxp3 suppression of class I expression in HeLa cells maps to the IRE and the core promoter.

Figure 4

A. Hela cells were cotransfected with WT class I promoter (2.0 µg) and Foxp3 (0 µg, 0.1 µg or 0.5 µg) or equal amounts of control vector. Results are expressed relative to class I promoter activity of the control co-transfection. Results are the average ± SEM of 3 experiments. ***p< 0.001.

B. Hela cells were cotransfected with 1.0 µg Foxp3 or control vector and 2.0 µg each of a series of 5’ truncation deletion mutants of the class I promoter, as diagrammed. The results for each construct are normalized to the control transfection with empty vector. Although the absolute activity of the constructs differed, all of them were transcriptionally active (12). The results are the average ± SEM of 4 separate experiments.

C. Hela cells were cotransfected with 1.0 µg Foxp3 or control vector and 2.0 µg each of a series 3’ truncation deletion mutants of the class I promoter. The results for each construct are normalized to the control transfection with empty vector. Although the absolute activity of the constructs differed, all of them were transcriptionally active. The results are the average ± SEM of 4 separate experiments.

D. Hela cells were cotransfected with 1.0 µg Foxp3 or control vector and 2.0 µg WT class I promoter or IRE mutation constructs. The results for each construct are normalized to the control transfection with empty vector. Although the absolute activity of the two constructs differed, both were transcriptionally active. The results are the average ± SEM of 4 separate experiments. **p=0.002.

E. HeLa cells were cotransfected with 1.0 µg Foxp3 or control vector and 2.0 µg of either WT or a MHC class I promoter construct containing a −50 to +3 bp deletion (drop-out). Although the absolute activity of the two constructs differed both were transcriptionally active. The results are the average ± SEM of 4 separate experiments. ***p<0.0001.

F. HeLa cells were cotransfected with 1.0 µg Foxp3 or control vector and 2.0 µg of either WT, IRE mutant, the drop-out or a mutant containing both the IREmut and the drop-out (IREm/drop-out). The results for each construct are normalized to the control transfection with empty vector. Although the absolute activity of the constructs differed, all of them were transcriptionally active (16).The results are the average ± SEM of 4 separate experiments. **p=0.004.