Fig. 5

ILF3 regulates ERp57 by promoting ERp57 mRNA stability. a SW839 cells transfected with shILF3 or A498 cells transfected with ILF3 overexpression vector pWP-ILF3. ERp57 mRNA level was measured by qRT-PCR. *P < 0.05 vs. corresponding control. b Cells were prepared as (a) and then exposed to Actinomycin D for 0, 2, 4, and 8 h. ERp57 mRNA level was detected using qRT-PCR. *P < 0.05, vs. corresponding control. c A498 cells were transfected with pWPI-ILF3 or empty vector. Western blot was used to measure the precipitation efficiency of ILF3 antibody. (d and e) RIP PCR and agarose gel electrophoresis was used to test the interaction between ILF3 protein and ERp57 mRNA. f A498 cells were transfected with ILF3-probe. Western blot analysis was used to test the ERp57, Cyclin D1, Cyclin E1 in pulldown precipitate. g SW839 cells were transfected with shERp57 or pWPI-ILF3 alone, or co-transfected with shERp57 and pWPI-ILF3. A498 cells were transfected with shILF3 or pWPI-ERp57 alone or co-transfected them together. Colony formation assay tested cell ability of proliferation. Right panel shows the analysis of colony formation number. *P < 0.05, ** P < 0.01 vs. corresponding control