Caspase-12 silencing altered NOD1, RIP2 and p-NF-κB protein expression in Leuk-1 cells.

<p><b>A</b> Immunofluorescence and confocal microscopy results showed that the transfection efficiency of Caspase-12 siRNAs was ∼95%. <b>B</b> FAM-labelled siRNAs indicated subcellular localization of cytoplasm, especially perinuclear area. <b>C</b> Western blot assay indicated protein level of Caspase-12 greatly decreased in Leuk-1 cells following RNA interference. <b>D</b> According to the image analysis result of immunoblot bands, compared with the NC group, the normalized protein level of Caspase-12 reduced remarkably in Caspase-12 siRNA-1 and -2 groups, especially for Caspase-12 siRNA-2. <b>E</b> Real-time PCR showed that the transfection of Caspase-12 siRNA-2 caused clear Caspase-12 silencing at mRNA level. <b>F</b> Western blot assay indicated Caspase-12 silencing altered NOD1, RIP2, and p-NF-κB protein expression. <b>G</b> Immunoblotting demonstrated that NOD1 expression significantly increased in Caspase-12-silenced cells compared with that in NC group. <b>H</b> Other than NOD1, Caspase-12 silencing remarkably reduced RIP2 level. <b>I</b> Alike to NOD1, p-NF-κB expression was up-regulated in Caspase-12-silenced cells. The immunoblot band density and mRNA data were expressed as means±SE (n = 3). Statistical significance: *<i>P</i><0.05, **<i>P</i><0.01, ***<i>P</i><0.001 vs. scrambled siRNA-transfected cells.