Fig. 1: Piwi and Gtsf1 interact with MEP-1, Mi-2, Rpd3 and p55 in OSCs. | Nature Communications

Fig. 1: Piwi and Gtsf1 interact with MEP-1, Mi-2, Rpd3 and p55 in OSCs.

From: The Mi-2 nucleosome remodeler and the Rpd3 histone deacetylase are involved in piRNA-guided heterochromatin formation

Fig. 1

a Western blot analysis of co-immunoprecipitation (co-IP) of Gtsf1-GFP from OSC nuclear extract using GFP-Trap. Control IPs were performed with OSCs transfected with a GFP-expressing vector. b Principle of dual-luciferase co-IP. A bait protein is fused to FLAG-tagged Firefly luciferase (FLAG-FFL), whereas potential interactors (preys) are fused to Renilla luciferase (RL). Following transient co-expression in S2R+ cells, the bait is immunoprecipitated with anti-FLAG antibodies and the co-IP efficiency is quantified by monitoring FFL and RL activities. c Graph shows the normalized co-IP efficiency (co-IP efficiency normalized to the RL-mCherry co-IP, this latter being used as a control of non-interacting prey) between the Gtsf1 bait and the indicated RL preys: Piwi, MEP-1, Mi-2, and Rpd3. Dots show values for n = 4 biologically independent samples, lines represent mean values ± SEM. d Western blot analysis of co-IP of endogenous Piwi and MEP-1 from Gtsf1-FH-containing OSC nuclear extract using anti-Piwi (left panel) and anti-MEP-1 (right panel) antibodies. Mouse and rabbit IgGs were used as control IPs for Piwi and MEP-1, respectively. The amount of input (In.) loaded relatively to IP is indicated as a percentage. Source data: c: Supplementary Data set 2; uncropped blot images are provided in Supplementary Data set 1.

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