Efg1p binds to the promoter of many Sfl1p and Sfl2p targets and co-immunoprecipitates with Sfl1p and Sfl2p, <i>in vivo</i>.

<p>(<b>A</b>) ChIP-PCR assay of selected Sfl1p and Sfl2p target promoters. Strains SFL1-TAP (CEC1922), SFL2-TAP (CEC1918) and EFG1-HA (HLCEEFG1) were grown in SC medium at 30°C (30°C) or in Lee's medium at 37°C (37°C) together with the SC5314 control strain (Control) during 4 h before being subjected to chromatin immunoprecipitation (Anti-TAP, Anti-HA) followed by PCR using primers specific to the indicated promoter regions. The <i>URA3</i> and <i>YAK1</i> genes were used as negative controls for ChIP enrichment. (<b>B</b>) Co-Immunoprecipitation of Efg1p with Sfl1p and Sfl2p. Strains coexpressing <i>SFL1-TAP</i> and <i>EFG1-HA</i> (Lanes 2 and 3) or <i>SFL2-TAP</i> and <i>EFG1-HA</i> (Lanes 7 and 8) or controls (Lanes 1 and 6, <i>EFG1-HA</i> only; lanes 4 and 9, <i>SFL1-TAP</i> only; lanes 5 and 10, <i>SFL2-TAP</i> only) were cultivated in SC medium at 30°C or in Lee's medium at 37°C before crosslinking with formaldehyde. Total extracts were incubated with Dynal PanMouse IgG beads directed against TAP epitope tag prior to washing and Western blotting using anti-TAP (IP Anti-TAP, 10% of the beads/total extracts mixture) and anti-HA (Co-IP Anti-HA) antibodies. A portion of the total cell extracts (∼2%) was included to verify the presence of the Efg1p-HA fusion (Total extracts Anti-HA).