Figure 5.

The intrinsic instability of the E12A-encoded protein product. A and B, E12A and EEF1A1 RNA abundance in different subcellular fractions of HeLa cells was determined by qPCR experiment (n = 4). Nuc, nucleus; Cyto, cytosol. C, ectopic expression vectors encoding WT HNRPLL (WT), unedited (A) and edited (G) versions of E12A were transfected into cells. Cells were treated with MG132 at the indicated time point. Immunoblotting analysis was performed to assess expression levels of the indicated proteins in the transfected and treated cells. D, expression levels of HNRPLL in the control and ADAR2-overexpressing cells were detected by immunoblotting. E and F, HeLa cells were transfected with empty vector, or expression vectors for FLAG-tagged WT HNRPLL, and N terminus- or C terminus-tagged E12A. Protein expression was monitored by Western blotting assay using the indicated antibodies. For immunoblotting data shown in this figure, β-actin expression was used as loading control.