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. 2005 Nov;25(21):9269–9282. doi: 10.1128/MCB.25.21.9269-9282.2005

FIG. 4.

FIG. 4.

Prp43p is associated with Pfa1p and with the RNA polymerase I machinery. Total extracts from cells expressing Pfa1p-TAP (lanes 1 and 2), Net1p-TAP (lanes 3 and 4), or Rpa190p-TAP (lanes 5 and 6) or devoid of tagged protein (lanes 7 and 8) were subjected to the tandem affinity purification procedure. In the case of the Pfa1p-TAP purification, 1/40th of the second fraction eluted from the calmodulin column was loaded in the immunoprecipitation (Ip) lane while in the case of the Net1p-TAP, Rpa190p-TAP, and control (No TAP-TAG) purifications, fractions eluted from the calmodulin column were pooled and all proteins present were precipitated using trichloroacetic acid and loaded in the Ip lanes. Tot: aliquot of the input extract. Proteins were subjected to SDS-PAGE, transferred to a cellulose membrane, and detected by enhanced chemiluminescence using affinity-purified anti-Prp43p polyclonal antibodies.