Figure 5.

Identification of the Pumpkin Phloem Proteins Coimmunoprecipitated with RBP50.

(A) Total phloem proteins (input protein) were coimmunoprecipitated using either preimmune serum or purified anti-RBP50 antibody (Ab). Proteins were visualized by GBS reagent. The co-IP complex(es) included interacting proteins IP1 to IP17; note that IP8 was confirmed to be RBP50 by LC-MS/MS analysis.

(B) Schematic illustration of phloem proteins that potentially could be bound to a phloem-mobile transcript contained within a RBP50–RNP complex. Note that transcripts bound by RBP50 may also interact with additional RNA binding proteins. RNase treatment was employed to separate a bound transcript (red darts) into individual RNP fragments, thereby allowing identification of proteins contained within the RBP50 complex.

(C) Co-IP of the RBP50 RNP complex was performed with or without RNase treatment. Asterisks indicate phloem proteins likely not directly contained within the RBP50 complex; darts indicate proteins likely bound to both the RBP50 and other protein complexes located on the same/different RNA molecule(s).