Skip to main content
. 2008 Dec 16;105(52):20852–20857. doi: 10.1073/pnas.0810958105

Fig. 4.

Fig. 4.

NPM/ALK induces CD274 expression through STAT3. (A) The effect of siRNA-mediated STAT3 and STAT5B depletion on CD274 mRNA expression. The ALK+TCL cell line SUDHL-1 was treated with siRNA specific for STAT3 or STAT5, STAT3/STAT5 siRNA combination, or control nonspecific siRNA and evaluated by RT-PCR for expression of mRNA coding for CD274 and the depicted other molecules serving as controls. (B) The effect of the siRNA-mediated STAT3 depletion on expression of the CD274 protein. SUDHL-1 cells treated with the STAT3, STAT5, STAT3/STAT5, or control siRNA were examined for CD274 protein expression by flow cytometry. (C) Binding of STAT3 to the CD274 gene promoter in vitro. The nuclear protein extracts from SUDHL-1 cells were incubated with the hot, biotin-labeled oligonucleotide probes corresponding to either of the two STAT3 binding sites identified within the CD274 gene promoter and analyzed in EMSA. The extract of the SUDHL-1 cells preincubated with the corresponding unlabeled cold probes served as control. (D) Binding of STAT3 to the CD274 gene promoter in vivo. Protein cell lysates from the SUDHL-1 cell line were analyzed in the ChIP assay using an anti-STAT3 rabbit polyclonal antibody and primer pairs specific for CD274 gene promoter. Non-immunoprecipitated lysates (input) and immunoprecipitates obtained with the STAT3 nonimmune entire IgG rabbit serum fraction served as a positive and negative control, respectively.