Fig. 1: The Erg11200-251 region and the Ncp11-63 region are important for the Erg11-Ncp1 interaction.
a The schematic diagram of a membrane yeast two-hybrid (MYTH) system. b MYTH analyzed the interaction between Erg11 and Ncp1. “Erg11” refers to yeast cells transformed with the bait Erg11 and the empty prey vector. “Erg11 + Ncp1” denotes the co-transformation of both the bait Erg11 and the prey Ncp1 into yeast cells. “Positive control” denotes the co-transformation of both the bait APP and the prey Fe65. “Negative control” refers to yeast cells that have been transformed with the bait APP and the prey vector without any protein insertions. c Co-IP assay analyzed the interaction between Erg11 and Ncp1. “Input” refers to lysates without any other manipulations used for immunoblotting. “IP” stands for immunoprecipitate, and the Protein A columns are used to collect the antibody-protein complex. “+“ indicates co-incubation of lysates with anti-Myc or anti-HA antibody, and “−“ represents lysates not incubated with antibodies. d Spotting assays of the yeast cells co-transformed to Ncp1 and C-terminal truncated fragments of Erg11. “Erg111-x “ represents the retained amino acids of Erg11. The schematic diagram depicts the interception strategy utilized to investigate the crucial interaction regions of Erg11. e Co-IP assay analyzed the interaction between Ncp1 and Erg111-251 or Erg111-200. The upper panel precipitates Erg111-251 with anti-Myc antibody and Ncp1 with anti-HA antibody. The lower panel precipitates Erg111-200 with anti-Myc antibody and Ncp1 with anti-HA antibody. f Spotting assays of the yeast cells containing Erg11 and N-terminal truncated fragments of Ncp1. “Ncp1y-680” represents the retained amino acids of Ncp1. g Co-IP assay analyzed the interaction between Erg11 and Ncp163-680 or Ncp191-680. The experiment was repeated three times with similar results. Source data, including uncropped and unprocessed scans with molecular weight markers are provided as Source Data file (c, e, g).
