PRL2 associates with and dephosphorylates CBL at tyrosine 371 in leukemia cells. (A) Knockdown of PRL2 decreased FLT3 half-life in MV-4-11 cells. (B) Knockdown of PRL2 enhanced FLT3 ubiquitination in MV-4-11 cells. (C) Total cellular proteins from MV-4-11 cells were isolated, incubated with GST, GST-PRL2, or GST- PRL2-CSDA and immunoblotted with antibody against FLT3, CBL, SHP2, and PLC-γ. (D) Co-immunoprecipitation assays showed that PRL2 interacts with FLT3 and CBL in MV-4-11 cells. (E) Immunofluorescence analysis showed that PRL2 colocalizes with CBL in MV-4-11 cells. (F) Representative western blot analysis showed that ectopic PRL2 expression decreases tyrosine phosphorylation of CBL in 293 cells. (G) Representative western blot analysis showed that knocking down of PRL2 increases the tyrosine phosphorylation of CBL in MV-4-11 cells. (H) Representative western blot analysis showed that knocking down of PRL2 increases CBL phosphorylation at tyrosine 371 in MV-4-11 cells. (I) Representative western blot analysis showed that ectopic expression of PRL2-CSDA increases CBL phosphorylation at tyrosine 371 in MV-4-11 cells. (J) APEX2-PRL2 proximity labeling was performed in HA-CBL or HA-CBL^Y371F transiently expressed 293 cells stably expressing APEX2-PRL2. After labeling, biotinylated proteins are enriched with neutravidin beads and examined with anti-HA and anti-PRL2 antibodies by western blot analysis. (K-L) PRL2-CSDA substrate trapping assays was performed in HA-CBL or HA- CBL^Y371F transiently expressed HeLa (K) or 293 (L) cells stably expressing Flag-PRL2-CSDA. After anti-FLAG pulldown, bound proteins were boiled in 50 μL Laemmli sample buffer and examined with anti-HA, anti-PRL2 antibodies by western blot analysis. CBL, Casitas B-lineage lymphoma.