The nuclear pore complex (NPC) is a cornerstone of eukaryotic cell functionality, orchestrating the essential transport of molecules between the nucleus and cytoplasm. Emerging evidence suggests its potential involvement in cell cycle regulation, but the specific roles and mechanisms of nucleoporins in this process remain largely uncharted. We previously found that PNET1 is a transmembrane nucleoporin and is expressed in actively dividing regions in Arabidopsis thaliana. Previous studies revealed that nucleoporins are phosphorylated during mitosis to facilitate the nuclear envelope breakdown process. In addition, we found that PNET1 extracted from callus tissues created a shifted band after running on Phos-tag gel, while this band was not observed when PNET1 is extracted from seedlings. We used LC-MS/MS to search the phosphorylation sites on PNET1 to further understand where exactly phosphorylation is happening and to create phospho-mimic and phospho-deficient versions of PNET1 later on. We detected more than 10 phosphorylation sites on PNET1.