Identification of C/EBPβ Target Genes in ALK+ Anaplastic Large Cell Lymphoma (ALCL) by Gene Expression Profiling and Chromatin Immunoprecipitation
Figure 5
Influence of G0S2 knockdown on cell proliferation, apoptosis and cell cycle.
Validation in primary cases. (A) Flow cytometric analysis of transduced SUDHL-1 cells and untreated controls four days after infection. The percentage of GFP-positive cells represents the percentage of infected cells. (B) RT-qPCR analysis of G0S2 mRNA in the transduced SUDHL-1 cells four days after infection. Values were normalized to TBP and data were analyzed according to the 2-ΔΔCp method. Results are depicted as mRNA amount relative to untreated SUDHL-1 cells. Error bars indicate SD (n = 3). (C) Proliferation curves of the controls and G0S2-shRNA infected SUDHL-1 cells up to 5 days after infection. Error bars indicate SD (n = 3). (D) Annexin V/propidium iodide staining of the controls and the G0S2-shRNA transduced SUDHL-1 cells four days after infection. (E) Cell cycle distribution of the control and G0S2-shRNA infected SUDHL-1 cells four days after infection. The differences between the uninfected control and the G0S2-shRNA-infected cells are given in percentage in the table. SUDHL-1 = uninfected cells, pG-NS = pGIPZ vector with non-silencing shRNA, pG-G0S2 = virus containing the pGIPZ vector and the G0S2-shRNA sequence, PI = propidium iodide. (F) Western Blot analysis of caspase 3 and cleaved caspase 3 in G0S2-shRNA transduced SUDHL-1 cells four days after infection. Each lane contains 30 µg cytoplasmic extract. 15 µg protein extract of Bortezomib-treated SUDHL-1 cells were used as control. α-Tubulin was used as loading control. (G) Box plot of G0S2 expression levels obtained by RT-qPCR analysis in 6 ALK+, 8 ALK- and 4 normal lymph node samples.