Deciphering NAC53, NAC78 interactome in response to Pst infection; NAC53, NAC78, MDP25 subcellular interactomes and identifying ubiquitination sites of NAC53 and NAC78 upon HRD1 E3 ligase mediated ubiquitination.

Analysis of NAC53 and NAC78 interactome in normal condition or upon Pseudomonas syringea pv. tomato DC3000 ∆HopQ infection. Constructs promUBQ10::eGFP-NAC53/eGFP-NAC78 are used as baits and promUBQ10::eGFP is used as control. Analysis of ubiquitination sites on NAC53 and NAC78 upon HRD1a/b ubiquitination. Analysis of NAC53, NAC78 and MDP25 subcellular interactome. For NAC53 and NAC78, dual tagged constructs pLjUBQ1::mCerrulean-NAC53/78-mCherry were used to pull-down N-ter moieties (cytosolic/nuclear localization) or C-ter moieties (endoplasmic reticulum localization). pUBQ10::eGFP and p35S::mCherry-HDEL are used as controls. For MDP25, protein variants were used, MDP25FL (plasma membrane localization), MDP25∆N25 (cytosolic localization) or N25MDP25 (plasma membrane localization). pUBQ10::eGFP is used as control.