Gene-expression changes in MM cells mediated by siRNA inhibition of FGFR3. (A) Western blot of FGFR3 protein expression in KMS11 cells at 48 hours following FGFR3-specific or scrambled control siRNA oligonucleotide transfection demonstrates successful knockdown of FGFR3. Nucleofections were done in KMS11 cells using the Amaxa apparatus and solutions. Transfection efficiencies were quantified using a green fluorescent protein (GFP) reporter construct provided by the supplier and flow cytometry. Our optimized nucleofection parameters yielded survival rates of 90% or more based on forward and side scatter parameters. Under these conditions, 70% of the surviving cells showed transgene expression 48 hours after nucleofection. (B) A heat map of triplicate experiments demonstrates differences in gene expression following FGFR3 knockdown, with representative examples of the most significant known dysregulated genes highlighted. A listing of 121 differentially regulated genes is provided in Supplemental Table S3.