Quantitative RT-PCR analyses of antisense transcripts after siRNA-mediated knockdown of antisense transcripts. Elevated levels of antisense transcripts seen in the Affymetrix antisense exon array analysis were confirmed by qRT-PCR for four antisense transcripts. These transcripts showed down-regulation after knockdown of antisense transcripts by specific siRNAs following transient transfection in a cell line. The levels of two control genes (marked by asterisks) unaffected by heat shock and devoid of Alus harboring HSF sites were monitored as experimental controls. These control genes, MGST1 and RPL8, did not show significant expression differences either during heat shock stress or after siRNA-mediated knockdown of the respective antisense transcripts. Error bars represent standard deviation among experimental replicates.
Pandey et al. Genome Biology 2011 12:R117 doi:10.1186/gb-2011-12-11-r117