Murine rack1 sirna

93T cells and L929 cells were grown in Dulbecco’s modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum, 100 U/ml penicillin and 100 μg/ml streptomycin. Plasmids or siRNAs were transfected into 293T cells with lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) according to the manufacture’s protocol, whereas transfection of L929 cells was performed by using Amaxa nucleofector II (kit V, Lonza, Switzerland) according to the manufacture’s protocol. RNA interference in primary murine hepatocytes was carried out with Accell siRNA (Dharmacon, Lafayette, CO, USA) utilizing the Accell siRNA delivery protocol. Plasmids used in this work have been described previously9 (link). Murine RACK1 siRNA (GGTCCAGGATGAGAGTCAT) and NC siRNA were from GenePharma (Shanghai, China).

Mammalian expression vectors encoding Myc-tagged murine HDAC1 and HA-tagged human Ube2T (MG53562-NM and HG124409-CY) were purchased from Sino Biological Inc. (Beijing, China). The mammalian expression vector encoding FLAG-tagged human RAB40C (CH816878) was obtained from Vigenebio (Jinan, Chian). Other mammalian or prokaryotic expression vectors used in this study have been described previously 13 (link), 19 (link). Murine RACK1 siRNA (GGGCAAGATCATTGTAGAT) and the non-targeting control siRNA were purchased from Shanghai GenePharma (Shanghai, China). Transfection was performed with Lipofectamine 2000 (11668019, Invitrogen, Carlsbad, CA, USA) according to the manufacturer's protocol.