Figure 1. JUNB depletion results in suppressed AKT1 mRNA and protein expression in ALK+ ALCL.
A. Karpas 299 cells were transiently transfected with increasing concentrations of JUNB siRNA, (+,50 nM and +++, 300 nM) or control (scramble, scr) siRNA which resulted in increasingly reduced JUNB (left) and AKT1 (right) mRNA levels as determined by qRT-PCR, which was performed in triplicate. B. AKT1 protein level was decreased following JUNB silencing, which was also associated with reduced levels of Ser473-phosphorylated (activated) AKT1 and reduced phosphorylation of its kinase substrate, GSK3. C. Similar results were obtained with another ALK+ ALCL cell line, SR-786. D. AKT1 protein and Thr308 phosphorylation levels were decreased in SR-786 cells following JUNB gene silencing. The expression level of JUNB and AKT1 mRNA was normalized to the GAPDH mRNA level (control) and it was shown as fold change to control.
*, p<0,05, **, p<0,01, ***, p<0,001 (Student’s t-test).