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. Author manuscript; available in PMC: 2016 Feb 24.
Published in final edited form as: Plant J. 2011 Jun 21;67(5):763–773. doi: 10.1111/j.1365-313X.2011.04630.x

Figure 1. Validation of the interactions between ZCCT1 and NF-Y proteins by in vitro and in planta Co-IPs.

Figure 1

GST antibody was used to immunoprecipitate protein complexes in the in vitro analysis, and Anti-HA Affinity Matrix was used to precipitate protein complexes in the in planta analysis. A&B) Western blot analyses of the in vitro Co-IPs, A) ZCCT1 antibody was used to detect the presence of ZCCT1 protein in the immunoprecipitates, B) the same membranes were stripped and re-hybridized with GST antibody. The proteins included in the in vitro assays are: 1) ZCCT1-MBP and GST tag; 2) ZCCT1-MBP protein alone; 3) ZCCT1-MBP and NF-YB1-GST; 4) ZCCT1-MBP and NF-YB9-GST; 5) ZCCT1-MBP and NF-YC11-GST; 6) ZCCT1-MBP and NF-YC7-GST; 7) ZCCT1-MBP and NF-YB3-GST; 8) ZCCT1-MBP and NF-YA7-GST; 9) ZCCT1-MBP and NF-YA9-GST; 10) ZCCT1-MBP and NF-YC10-GST. C&D) Western blot analyses of the in planta Co-IPs, C) Anti-Myc antibody was used to detect Myc-NF-YB fusions, D) the same membrane were stripped and re-hybridized with Anti-HA antibody. Protein extracts of the following infiltrations were used in the in planta Co-IPs: 1) Infiltration of HA-ZCCT1 alone; 2) Co-infiltration of HA-ZCCT1 and Myc-NF-YB3; 3) Co-infiltration of HA-ZCCT1 and Myc-NF-YB1; 4) Co-infiltration of HA-ZCCT1 and Myc-NF-YB9.