Supplementary Figure 3: Depletion of SETDB1 decreases Akt kinase activity.

a-c, IB analysis of GST pull-down (a,b) or IP (c) products and WCL derived from HEK293T cells transfected with indicated constructs. d, IB analysis of cell fractionations separated from HEK293, DLD1 and A375 cells, respectively. e-g, IB analysis of dot blot (e) and IP products derived from HEK293 cells transfected with the indicated constructs (f,g). h, In vitro methylation assays were performed with IP Flag-SETDB1 derived from HEK293T cells as the source of methyltransferase, and the commercially purified His-Akt1 as the substrate. i-k, IB analysis of IP products, GST pull-down and WCL derived from HBL cells lentivirally infected with shRNAs against SETDB1 (i), or derived from HEK293 cells transfected with the indicated constructs (j,k). l-o, IB analysis of Setdb1 conditional knockout MEFs treated with or without 4-OHT (500 nM) in different time points (l,m). Resulting cells were subjected to colony formation assays (n). The experiment was performed twice independently with three repeats, and exhibit similar results (n). Representative images were shown in n and relative colony numbers derived from two independent experiments were plotted in o. Source data for o are shown in Supplementary Table 2. All Western-blots above were performed twice independently with similar results. Scanned images of unprocessed blots are shown in Supplementary Fig. 8.