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Fig. 2 | BMC Molecular Biology

Fig. 2

From: Sumoylation in p27kip1 via RanBP2 promotes cancer cell growth in cholangiocarcinoma cell line QBC939

Fig. 2

RanBP2 can increase the sumoylation of p27kip1. RanBP2-HA plasmid was constructed and transfected to QBC939 cells. Immuno-precipitation with anti-p27kip1 antibody verified that RanBP2 can increase the sumoylation of p27kip1 (a). Also, flag-p27kip1 plasmid was constructed and transfected into either QBC939-RanBP2-HA or QBC939-control cells. Immuno-precipitation with anti-HA antibody proved that p27kip1 can bind with RanBP2 (b). Correspondingly, HA-RanBP2 plasmid was transfected into either QBC939-flag-p27kip1 or QBC939-control cells. Immunoprecipitation result with anti-flag antibody proved that RanBP2 can bind with p27kip1. c. Moreover, GST-RanBP2 and His-p27kip1 plasmids were constructed for GST-pull down assay. As shown in d, p27kip1 proved to be one of the binding targets of RanBP2. e HA-RanBP2 plasmids and its control vector were transfected into QBC939 cells. Cytoplasmic proteins, nuclear proteins and total cellular proteins were separately extracted from the same batch of cells and sent for WB. Also, RanBP2 was validated to be overexpressed after HA-RanBP2 transfection. f In vitro sumoylation assay was undertaken by his-tagged p27kip1 proteins were incubated with SAE1/SAE2, SUMO1 and UBC9 in increasing doses (**p < 0.01)

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