Supplementary Figure 1. Densitometoric Analysis of Immunoblotting. A, Ratios of P-4E-BP1

Supplementary Figure 1.Densitometoric analysis of immunoblotting. A, Ratios of P-4E-BP1 to 4E-BP1, P-p70 S6K to p70 S6 K, and P-Akt to Akt were measured, and values in untreated (control) cells were set as 1. B, Ratio of cleaved PARP to PARP was measured, and the value in control cells was assigned as 1. C, Ratio of LC3II to LC3I was measured, and the value of control cells was assigned as 1.

Supplementary Figure 2.Rapamycin inhibits proliferation of T and NK cell lines. A,Jurkat, SNT13, KHYG1 and KAI3 were treated with the indicated concentrations of rapamycin and viable cells were counted using the trypan blue exclusion test. Values are means ± SE of results performed in four replicates. *, P <0.05 as compared with controls. B, cell proliferation was measured by MTS assay. Cells were incubated with the indicated concentrations of rapamycin for 3 days. Values are means ± SE of results from triplicate experiments. *, P <0.05 as compared with controls. C, T and NK cell lines were treated with 25 nMrapamycin for 2-6 days. Cell viability determined by trypan blue exclusion test is shown as the ratio of viability in different treatment groups to DMSO-treated cells. Values are means ± SE of results from triplicate experiments.

Supplementary Figure 3.CCI-779 induces inhibition of cell proliferation by G1 cell cycle arrest in Epstein-Barr virus (EBV)-positive T and NK cell lines. A, AnEBV-positive T cell line (SNT16) and an EBV-positive NK cell line (SNK6) were treated with the indicated concentrations of CCI-779 for 72 hours. Cell number is shown as the ratio of cell numbers in the different treatment groups to ethanol-treated cells. Values are means ± SE of the results from triplicate experiments. B, SNT16 and SNK6 cell lines were treated with 10 nM CCI-779 for 1 hour (P-4E-BP1 and P-p70 S6K) or 24 hours (p27, CDK2, Rb and β-actin), and lysates were blotted for the proteins indicated. C, SNT16 and SNK6 cell lines were treated with the indicated concentrations of CCI-779 for 24 hours, and were then fixed and stained with propidium iodide. Cell cycle profiles were assessed by flow cytometry.*, P <0.05 as compared with controls.

Supplementary Figure 4.CCI-779 inhibits tumor growth and proliferation in the murine xenograft model. CCI-779 inhibits growth of subcutaneous xenograft tumors in NOG mice. SNK6 cells (1×106 cells/flank) were subcutaneously inoculated into the flanks of mice. Four days after inoculation of SNK6 cells, mice were treated with CCI-779 (10 mg/kg, i.p.) for three weeks, and tumor size was quantified twice weekly. *, P<0.05; n = 6 mice for each group.