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. 2008 Feb 15;9(3):280–286. doi: 10.1038/embor.2008.10

Figure 2.

Figure 2

Lack of NEDDylation decreases the levels of L11 ribosomal protein. (A) Flag-L11 and NEDP1 were expressed in H1299 cells, as indicated. Cells were lysed in 2 × SDS sample buffer and equal amounts of protein were analysed by western blotting. Transfection efficiency and loading were monitored by β-gal antibodies. (B) The experiment was performed as above using the SENP6 expression construct. Expression of NEDP1 and SENP6 was monitored with NEDP1 or SV5 antibodies, respectively. (C) TS-41 or the parental Chinese hamster ovary (CHO) cells were grown at 32°C. Fifteen hours before cells were collected, some cells were transferred to 39°C. Extracts prepared in 2 × SDS sample buffer were analysed for endogenous L11 ribosomal protein. (D) H1299 cells were transfected with 20 nM of the indicated siRNA. At 48 h after transfection, cells were lysed in 2 × SDS sample buffer and equal amounts of protein were analysed by western blotting. Endog., endogenous; NAE1/APPBP1, NEDD8-activating enzyme E1 subunit 1; siRNA, short interfering RNA.