Effect of IRE1a and XBP1 knockdown on gene expression in primary mouse keratinocytes expressing an HRas oncogene

IRE1a is a critical modulator of the unfolded protein response. Its RNAse activity generates the mature transcript for the XBP1 transcription factor and also degrades other ER associated mRNAs in a process termed Regulated IRE1a Dependent mRNA Decay or RIDD. To determine if IRE1a is critical in the response to oncogenic Ras we used ShRNA to knockdown Ire1a or Xbp1 in primary mouse epidermal keratinocytes transduced with a v-HRAS retrovirus. Primary mouse keratinocytes were transduced with a v-Hras retrovirus and either a control ShRNA lentivirus or ShRNA against either Ire1a or Xbp1. RNA isolated from each group at 4 days post lentiviral infection (n=3 biological replicates per group) was used for microarrays. Primary mouse keratinocytes transduced with control ShRNA lentivirus served as the reference set for gene expression analysis (n=3 biological replicates per group).