PKA-activated ApAF-ApC/EBP heterodimer is a key downstream effector of ApCREB and is necessary and sufficient for the consolidation of long-term facilitation -- Data Supplement - JCB--Lee et al. -- The Journal of Cell Biology

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Figure S2. Interaction of ApAF with ApC/EBP and ApCREB2. Interaction of translated ApCREB2-HA and ApC/EBP-HA proteins with bacterially expressed glutathione S-transferase fusions of full-length WT ApAF (GST-ApAF [WT]), ApAF mutant (GST-ApAF [S266A]), and ELAV1 (negative control). Glutathione-agarose resins saturated with GST fusion proteins were incubated with 5 µl of each translated HA fusion protein, washed, eluted, and detected with anti-HA rabbit antibody. Both ApCREB2-HA and ApC/EBP-HA form heterodimers with WT ApAF or ApAF mutant, but not with the RNA-binding protein ELAV1. Both ApCREB2 and ApC/EBP interact with ApAF(S266A) mutant with an affinity comparable to that of WT ApAF(WT).