Comprehensive Dissection of PDGF-PDGFR Signaling Pathways in PDGFR Genetically Defined Cells
Figure 2
Representative PDGFR dependent differential gene expression in response to PDGF treatment.
Upper panel: triplicate sets of each cell line were treated with or without PDGF-BB for 1 h. mRNA expression profiles for the probe sets identified as differentially expressed in the treatment group for all of the cell lines, except for double knockout. Expression levels of the probe sets interrogating the genes that were commonly differentially expressed in the PDGFRβ−/− (beta null), PDGFRα−/− (alpha null), and WT (PDGFRα+/+ PDGFRβ+/+) cell lines are shown in the heatmap. Each column represents a sample, each row a gene. Column labels indicate the cell line and treatment condition (T for samples treated with PDGF-BB, U for untreated samples) for each sample. Each probe set's expression has been independently normalized across the experiments. Bright green shading indicates an expression level below the gene-wise mean, bright red indicates an expression level above the mean, while darker shades indicate expression levels closer to the mean intensity. Lower panel: four cell lines were treated with PDGF-BB (50 ng/ml) for various times and their lysates were immunoblotted for ATF-3, Txnip, Fra-1, Nurr1, Nur77, TF, and EGR1 (PDGF treatment does not influence the total Erk, therefore Erk was regarded as a spotting control in this system, the same for the following experiments in the MEF cell lines with genetically defined PDGFRs) to validate findings from the mRNA expression analysis (upper panel). Expression levels of probe sets interrogating the genes for these proteins are shown in the heatmap above the western blot figure.