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. 2000 Mar 7;97(7):3118–3123. doi: 10.1073/pnas.050007597

Figure 4.

Figure 4

The ubiquitylation status of GAL4-VN8 proteins is directly related to transcriptional potency. HeLa cells were transfected with expression constructs encoding the indicated GAL4-fusion proteins, either alone (odd-numbered lanes) or with an expression construct encoding polyhistidine-tagged human Ub (His-Ub; even-numbered lanes). After transfection, cells were treated with LLnL to allow the unstable GAL4-derivatives to accumulate. Cellular proteins were harvested as described in Materials and Methods, and HA-tagged GAL4-fusion proteins present in the total lysate (lanes 1–10) or the nickel-affinity-purified material (lanes 11–20) were revealed by SDS/PAGE and immunoblotting. The arrows indicate the position of His-Ub-GAL4–6×VN8 conjugates (lane 18).