Fig. 8
OsBAG6 interacts with OsCaM1-1 both in vivo and in vitro. A Venn diagram showing overlap of proteins identified by affinity purification and mass spectrometry analysis in two repeats. B OsBAG6 binding proteins identified by IP-MS. The percentage of full-length protein covered by identified unique peptides were defined as Coverage. Unique peptides indicate the number of identified peptides that are mapped to an individual protein. C GO enrichment analysis of 276 genes shade in red encoding OsBAG6 and OsBAG6 interacting proteins in Fig. 8A. D Immunoblot analysis of the results from the co-immunoprecipitation (co-IP) assay. OsBAG6-FLAG construct was co-transfected with OsCaM1-1-HA into Kitaake protoplasts. OsBAG6-FLAG and OsCaM1-1-HA were separately transfected as negative control. Anti-FLAG antibody was used to perform immunoprecipitation, and pulled-down proteins were detected using anti-HA antibody. E Yeast two hybrid (Y2H) assay was performed to test the binding between OsBAG6 with OsCaM1-1 in vitro. F Pull-down assay was used to test the binding affinity between OsBAG6 with OsCaM1-1 in the presence of 5 mM EGTA or 2 mM Ca2+. Magnetic beads attaching anti-FLAG antibody was used for pull-down