Fig. 2: Interaction between NOTCH1 intracellular domain (NICD1) and mitochondrial pyruvate dehydrogenase E1 subunit beta (PDHB). | Nature Communications

Fig. 2: Interaction between NOTCH1 intracellular domain (NICD1) and mitochondrial pyruvate dehydrogenase E1 subunit beta (PDHB).

From: NOTCH1 mitochondria localization during heart development promotes mitochondrial metabolism and the endothelial-to-mesenchymal transition in mice

Fig. 2

A Co-immunoprecipitation (CO-IP) of exogenous NICD1 and PDHB in human embryonic kidney 293T (HEK293T) cells. B, CO-IP of endogenous NICD1 and PDHB in rat cardiac endothelial cells. C Left: Schematic of truncated NICD1. RAM, Rbp-associated molecule domain; ANKs, ankyrin repeats; TAD, transcription activation domain; PEST, proline (P), glutamic acid (E), serine (S), threonine (T) degradation domain; NLS, nuclear localization signals. Right: CO-IP of exogeneous truncated NICD1 and PDHB in HEK293T cells. D Left: Schematic of recombinant proteins constructed for bimolecular fluorescence complementation (BiFC) assays: PDHB fused with N-terminus of Venus (VN-PDHB) and NICD1 fused with C-terminus of Venus (VC-NICD1) in reaction 1, NICD1 fused with VN (VN-NICD1) and PDHB fused with VC (VC-PDHB) in reaction 2, and bJun (the binding domain of Jun [the 257–318 amino acids]) fused with VN (VN-bJun) and bFos (the binding domain of Fos [the 118–210 amino acids]) fused with VC (VC-bFos) in positive control. Right: Representative western blot illustrating the expression level of recombinant proteins in HEK293T cells. E–G BiFC assay exhibiting the interactions of VN-PDHB and VC-NICD1 in reaction 1(E), VC-PDHB and VN-NICD1 in reaction 2 (F), and VN-bJun and VC-bFos in positive control (G) in green dots and their intracellular distribution (Scale bar = 20 μm). Three experiments were repeated independently with similar results for A–G. Source data are provided as a Source Data file.

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