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. 2000 Feb 15;28(4):952–959. doi: 10.1093/nar/28.4.952

Figure 2.

Figure 2

Coexpression of various combinations of the subunits and formation of subunit complexes. (A) Coexpression of Pol II subunits. The complete set of 11 Pol II subunits or various combinations of 10 subunits were expressed by mixed infection of the recombinant viruses. Lysates of the infected cells were subjected to western blotting and detected with antisera against each subunit. Portions of the filters corresponding to the migration positions of each subunit in the SDS–PAGE are arrayed, and the detected subunits are indicated on the left. The proteins expressed in each lane are: GUS, β-glucuronidase; –Rpb3/+GST, GST plus 10 subunits except Rpb3; 11 subunits, 11 subunits including GST–Rpb3; –Rpb1H to –Rpb12, 10 subunits (including the GST–Rpb3) except for the indicated subunit. (B) Isolation of subunit complexes. The cell extracts were subjected to GSH-affinity chromatography. The eluted fractions were analyzed by western blotting as in (A). Lanes correspond to those in (A) with the same designations. (C) Preferential assembly of the intact Rpb1H. Western blotting with anti-Rpb1 antiserum was carried out: GUS/lysate, the lysate of cells expressing GUS; 11 subunits/lysate, the lysate of cells expressing the 11 subunits; 11 subunits/GSH-affinity, the subunit complex fraction isolated by GSH-affinity chromatography.