Supplementary Figure 3: Expression of control baits and autoactivation assay. | Nature Methods

Supplementary Figure 3: Expression of control baits and autoactivation assay.

From: Post-translational selective intracellular silencing of acetylated proteins with de novo selected intrabodies

Supplementary Figure 3

(a) LexA control baits used for epitope mapping are expressed in yeast to investigate their correct expression, as done for the screening baits. The only bait that does not carry a HA tag is LexA-Integrase, whose expression is shown, in the central figure, for two different clones (CFUs). (b) Auto-activation assay for screening and control baits. Yeast baits spotted on histidine-lacking plates in which 3-AT is titrated, allow identification of auto-activation level. The 3-AT concentration, at which non-specific growth (in the absence of an interactor) is arrested, is included in the medium when testing an intrabody interaction. H3 screening bait (LexA-H3-Gcn5wt-HA) needs at least 5mM 3AT, while other control baits are grown at 1.25mM 3AT. As for the Integrase bait panel, LexA-IN-P300wt-HA does not show any auto-activation, unlike LexA-P300wt-HA and LexA-IN which require 1.25mM 3AT.

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