UCH-L1 promotes AKT phosphorylation and cell survival in DLBCL. (A) The expression of UCHL1 is shown for a series of GCB-DLBCL cell lines. The cell lines used in this study are shown in red (UCHL1 HI) and green (UCHL1 LO). Data extracted from the CCLE (GSE36133) and analyzed using the R2: Genomics Analysis and Visualization Platform (http://r2.amc.nl). (B) Immunoblot analysis of the cell lines from panel A. Histone H2B is included as a loading control. (C-D) The cell lines from panel A were transduced with lentivirus’ encoding the indicated Dox-inducible shRNA constructs. Cell viability (C) was monitored using the MTS assay. The level of UCH-L1 (D) is shown for the WSU-CLCL2 cell line. shNS, control nonsilencing; shUCHL1, n = 3 independent UCHL1 targeting shRNA constructs. Similar results were obtained in SU-DHL6 cells. (E) The indicated cell lines were transduced with the indicated shRNA constructs as in panels C and D and the resulting extracts were analyzed with antibodies against the indicated proteins. Dox was included where indicated.