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. 2012 Aug 3;31(17):3596–3606. doi: 10.1038/emboj.2012.218

Figure 2.

Figure 2

A targeted siRNA screen identifies a role for MEX-3C, a novel RNA-binding E3 ubiquitin ligase, in HLA-A2 regulation. (A) siRNA-mediated depletion of MEX-3C significantly increases cell-surface HLA-A2 expression. The left bar chart shows the Z-scores for the 375 E3 ligases screened in three independent experiments. Of the 375 E3 ligases screened, only MEX-3C depletion gave a significant increase in HLA-A2 expression. The graph on the right shows the top five Z-scores. MEX-3C’s Z-score (5.23) was significantly above the Bonferroni-corrected P-value threshold (P=0.05) of 3.82 (dashed line). (B) Flow cytometric analysis of cell-surface HLA-A2 levels in mock (dashed line) versus siRNA-treated cells (solid line). Isotype control (solid grey). (C) Stable shRNA-mediated depletion of MEX-3C increases cell-surface HLA-A2. (Left) Cytofluorometric analysis of HLA-A2 surface levels in parental (mock shRNA depleted, dashed line) HEK293 and cells stably transduced with a shRNAmir against MEX-3C (shMEX-3C; solid line). (Right) Immunoblot analysis of MEX-3C in HEK293 cells expressing myc-tagged MEX-3C or control cells (mock), both stably transduced with shMEX-3C. The shMEX-3C sequence is identical to the targeting oligonucleotide #2 of the original screening pool. β-Actin was used as loading reference levels.