Fig. 6

USP37 stabilizes PCNA (Proliferating Cell Nuclear Antigen) with and without replication stress and interacts with PCNA. A Cells were treated with Hydroxyurea (100 μM) and cycloheximide (100 μg/ml) for up to 4 h and then harvested to perform WB with Anti PCNA antibody. B Cells were transfected with Myc USP37 and treated with Hydroxyurea (100 μM) and cycloheximide (100 μg/ml) for upto 4 h and then harvested to perform WB with Anti PCNA antibody. C. Cells were treated with HU (100 μM) and Cisplatin (75 μM) for 24 h and then harvested to perform Immunoprecipitation. IP was done using an Anti-USP37 antibody while WB was done using an Anti PCNA antibody. D Cells were treated with HU (100 μM) and Cisplatin (75 μM) for 24 h and then harvested to perform Immunoprecipitation. IP was done using Anti PCNA antibody while WB was done using Anti USP37 antibody. E Cells were transfected with Myc USP37 and treated with HU (100 μM) and Cisplatin (75 μM) for 24 h and then harvested to perform Immunoprecipitation. To check the interaction of PCNA with USP37 IP was done using Anti PCNA antibody while WB was done using an anti-Myc antibody F Cells were transfected with Myc USP37 and treated with HU (100 μM) and Cisplatin (75 μM) for 24 h and then harvested to perform Immunoprecipitation. Reverse IP was done and capture was done using Anti Myc antibody while WB was done using anti-PCNA antibody. G. U20S cells were transfected with plasmids encoding HA-Ub, HA-Ub + myc USP37, HA-Ub + USP37 350A and treated with 100 μM hydroxyurea (HU) and 10 μg/ml MG132 as indicated. Lysates and ubiquitin-conjugated proteins were analyzed by immunoblotting as indicated. H. U20S cells were transfected with plasmids encoding HA-Ub, HA-Ub + myc USP37, HA-Ub + USP37 350A and treated with 100 μM hydroxyurea (HU) and 10 μg/ml MG132. Lysates and ubiquitin-conjugated proteins were analyzed by immunoblotting by respective antibodies as indicated