Skip to main content
. Author manuscript; available in PMC: 2013 May 1.
Published in final edited form as: Cancer Res. 2012 Mar 15;72(9):2251–2261. doi: 10.1158/0008-5472.CAN-11-3386

Figure 2. Transcriptional regulation of the SASP factors OPN versus IL6 and IL8 is governed by distinct mechanisms.

Figure 2

A. NF-κB activity was examined in BJ fibroblasts stably expressing a vector control or IκBα-mut allele. Cells were transiently transfected with an NF-κB-responsive promoter driving Firefly luciferase. Data are presented as percent Renilla Luciferase, a control for transfection efficiency. The mean ± SEM is shown (n=3). B. SA-βgal staining of BJ fibroblasts stably transduced with a vector control or IκBα-mut allele and treated with vehicle (Vector+Vehicle) or bleomycin (Vector+Bleo and IκBα-mut+Bleo). Scale bar is 100 microns. C. The mRNA levels of OPN, IL6, and IL8 as indicated were measured by qPCR in vector control BJ fibroblasts treated with vehicle (Vector) (set to 1) or bleomycin (Vector+Bleo) or BJ fibroblasts expressing the IκBα-mut cDNA and treated with bleomycin (IκBα-mut+Bleo). The mean ± STDEV is shown (n=3). D. ATM mRNA levels were quantified by qPCR in BJ fibroblasts expressing one of two short-hairpin RNAi constructs targeting ATM (shATM-1 or shATM-2) or a control hairpin (shSCR), which was defined as 100 percent. The mean ± STDEV is shown (n=3). E. SA-βgal staining of control hairpin cells treated with vehicle alone (shSCR+Vehicle) or bleomycin (shSCR+Bleo) and shATM expressing BJ fibroblasts treated with bleomycin (shATM-1+Bleo). Scale bar is 100 microns. F. The mRNA levels of OPN, IL6, and IL8 were measured by qPCR in BJ fibroblasts expressing a control hairpin treated with vehicle or bleomycin (shSCR+Vehicle and shSCR+Bleo) or a short-hairpin targeting ATM and treated with bleomycin (shATM-1+Bleo and shATM-2+Bleo, respectively). Expression levels in shSCR+Vehicle cells were set to 1 for each factor measured. The mean ± STDEV is shown (n=3).