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. Author manuscript; available in PMC: 2019 Jul 12.
Published in final edited form as: Structure. 2017 Sep 14;25(10):1519–1529.e4. doi: 10.1016/j.str.2017.08.002

Figure 4. Biochemical Verification of the Importance of Insertion Domain in ScR2TP Assembly.

Figure 4.

Solid ovals represent Rvb1p (1), Rvb2p, or Rvb2p DII deletion (2), Pih1p (P), and Tah1p (T). SDS-PAGE gels are stained by either Coomassie brilliant blue (blue) or silver nitrate (yellow). Samples analyzed on gels are denoted as L for load, B for bound, PT for Pih1p-Tah1p control, and M for protein markers.

(A) Schematic representation of the assembly and pull-down method (left) and SDS-PAGE gel analysis of Pih1p-Tah1p binding to Rvb1/2p lacking domain II in Rvb2p (ScR2TP-ΔII2) or DII2 (domain II of Rvb2p) (right).

(B) SDS-PAGE gel (silver-stained), showing Pih1p-Tah1p binding to Rvb2p (R22TP) but not to Rvb2p lacking its domain II (R22TP-ΔII). The last lane (R12-ΔII2) shows binding of Rvb1p to the same Rvb2p lacking its domain II (Rvb2p-ΔII) sample used for the R22TP-ΔII complex formation.

(C) SDS-PAGE gel, showing formation of the Pih1p-Tah1p-Rvb1p complex (R1TP).