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. Author manuscript; available in PMC: 2014 Nov 15.
Published in final edited form as: J Immunol. 2013 Oct 14;191(10):10.4049/jimmunol.1302079. doi: 10.4049/jimmunol.1302079

Figure 1. Genomic siRNA library screening strategy and representative images of immunofluorescence analysis.

Figure 1

(A) siGenome siRNA library (Dharmacon) screening testing involved transfecting PC3 cells with individual SMARTPools containing 4 distinct siRNAs targeting a single mRNA and arranged in 96-well plates format. Validation screen tested individual siRNAs (Dharmacon) from the corresponding SmartPool “hits” and arranged in 96-well plates format. (B) As an internal negative control, cells were introduced with SMARTPool siRNA targeting luciferase (siGL2). As a positive control, siRNA silencing with β2m specific siRNA SMARTPool significantly reduced β2m surface expression on PC3 cells (siβ2m) as compared to siGL2 control. Cells were stained with anti-β2m mAb and APC-conjugated secondary Ab. (C) siRNA silencing with SH3ST3B1 specific siRNA SMARTPool significantly reduced KIR2DL4 ligand(s) surface expression on PC3 cells as compared to siGL2 control. Cells were stained with KIR2DL4-Ig and APC-conjugated secondary Ab.