Additional file 3.
Supplemental figure S1. (A) Validation of CSMA transfection efficacy in VCaP cells was performed using four different siRNA constructs for CAPN2 and a control siRNA. Composite (CAPN2 = red, DNA = blue) and single channel confocal laser microarray scanned images of a VCaP array with four 24 replicate spot sub-grids of three CAPN2 and control siRNA constructs. Scale bar 900 μm. (B) 20× microscopic images of control siRNA and CAPN2 siRNA spots with an intensity surface plot visualization of the intra-spot signal distribution of anti-CAPN2 staining (lower panels). (C) Upper panel: On basis of the immunofluorescent detection of CAPN2 an up to 94% spot level efficacy was measured with spot level normalization against DNA counterstaining of the cells. The mean efficacy of CAPN2 siRNAs varied from 31% to 94% silencing. Lower panel: Comparative Western blot analysis of cells transfected using conventional methods provided identical efficacy profiles for the siRNA constructs. (D) Western blot of analysis of cells recovered from a CSMA array of 384 replicate CAPN2 and control siRNA spots after 72 h transfection was used to validate the transfection efficacy in VCaP and PC-3 prostate cancer cells and SVpgC2a oral keratinocytes.
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Rantala et al. BMC Genomics 2011 12:162 doi:10.1186/1471-2164-12-162