FIG. 5.

AIRE recruits P-TEFb to Ins2 and Spt1 promoters and stimulates transcriptional elongation by RNAPII. 1C6 (−AIRE) and 1C6.AIRE (+AIRE) cells were analyzed. Formaldehyde-fixed and sonicated chromatin extracts were immunoprecipitated with the indicated antibodies. ChIP/qPCR was performed with the indicated primers to determine the amounts of DNA associated with immunoprecipitated proteins on promoters or coding sequences. The positions of primers are indicated in the diagrams above the graphs. We looked for the presence of AIRE, RNAPII, and subunits of P-TEFb on AIRE-responsive genes (A and B) and AIRE-nonresponsive genes (C and D). ChIP/qPCR with rabbit IgG antibodies was used as the negative control for specificity. All values are expressed relative to the control input DNA (% input) and represent experiments performed in triplicate, with errors indicated.