Fig. 9.
OsSEC3A physically interacts with OsSNAP32. (A) Y2H assay showing that full-length OsSEC3A and OsSEC3A_C (amino acids 521–888) interact with OsSNAP32, while two other predicted domains, OsSEC3A1–160 (including the PH domain) and OsSEC3A190–510 (including Vps52) failed to interact with OsSNAP32. (B) In vitro pull-down assay of recombinant MBP–OsSEC3A, MBP–OsSEC3AC (amino acids 310–888, C-terminal half), and MBP using resins containing GST–OsSNAP32. Asterisks indicate MBP–OsSEC3A and MBP–OsSEC3AC proteins. (C) Quantification of luciferase (LUC) activity in N. benthamiana leaves expressing OsSEC3A_C-NLuc and CLuc-OsSNAP32. The western blot shows the expression levels of CLuc and NLuc fusion proteins. OsSEC3A_C derivatives were detected by anti-firefly LUC antibodies, whereas CLuc-OsSNAP32 was detected by anti-His antibodies. The staining shows equal loading of protein in each lane. Data were collected 3 d after infiltration. The data shown are representative of three independent experiments.