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Fig. 5 | Epigenetics & Chromatin

Fig. 5

From: Eviction of linker histone H1 by NAP-family histone chaperones enhances activated transcription

Fig. 5

SET enhances activated transcription in vivo. a SET, but not hNAP1, enhanced activated transcription from the chromosomally-integrated model promoter. CHOK-Luc cells, carrying multiple integrated copies of the HTLV-1 promoter linked to luciferase [63] were transiently transfected with an expression plasmid for Tax alone (10 ng) or co-transfected with increasing amounts (25, 50, 100, 200, 400, 500 ng) of HA-hNAP1 or HA-SET expression plasmids. The signal obtained in the presence of Tax alone was normalized to 1, and fold activation by the histone chaperones, relative to Tax alone, is reported. The experiment was performed in triplicate, and is representative of three biological replicates. b Expression of HA-hNAP1 (upper panel) and HA-SET (lower panel) following transfection. CHOK-Luc cells were transfected as described in panel a, and lysates analyzed by immunoblot using an anti-HA antibody. Immunoblot analysis of GAPDH was performed in parallel as a loading control. c SET enhancement of transcription requires co-transfection of Tax. CHOK-Luc cells were transiently transfected with an expression plasmid for Tax (10 ng) or the expression plasmid for SET (200, 400, 500 ng), as indicated. d The transiently transfected HTLV-1 promoter/luciferase reporter plasmid is refractory to enhancement by co-transfected SET. Jurkat T-cells were transiently co-transfected with the HTLV-1/Luc reporter plasmid (100 ng) [87], and expression plasmids for Tax and SET, as described in panel a. e SET/Taf1α, an isoform of SET/Taf1β, enhanced activated transcription from the integrated model promoter. The experiment was performed as described in panel a, with co-transfected Taf1α in place of SET. f Co-transfection of hNAP2 had a negligible effect on activated transcription from the integrated model promoter. The experiment was performed as described in panel a

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