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. 2016 Aug 9;7(37):59519–59534. doi: 10.18632/oncotarget.11148

Figure 3. The depletion of U3 or U8 elicits a p53-dependent nucleolar antitumor surveillance pathway.

Figure 3

(A) U3 and U8 depletion lead to an increased p53 steady-state level. Total protein extracted from H1944 or MCF-7 cells depleted of U3 or U8 for 1, 2, or 3 days was resolved on SDS-polyacrylamide gels and analyzed by Western blotting with specific antibodies (see Materials and Methods). As a control, cells were treated with a non-targeting silencer (SCR). In H1944 cells, at the late time points of depletion (48 and 72 h), p21 was detected as a doublet, suggesting it is post-translationally modified. The signals were quantitated with a ChemiDoc and normalized with respect to SCR-treated cells. As loading control, we probed the blots for β-actin. (B) The increase in p53 steady-state level observed upon U3 or U8 depletion depends on the presence of uL5 and uL18. H1944 and MCF-7 cells depleted of U3 or U8, or treated with a non-targeting control silencer (SCR), were codepleted of uL5 or uL18. Total protein was extracted and analyzed as in panel A. uL5 appears as a doublet, suggesting it is post-translationally modified.