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This study explores the effects of various small interfering RNAs (siRNAs) on apoptosis-related proteins, including Caspase-2 and Caspase-9, in HuT102 cells. We analyzed the relative mRNA expression levels of RAIDD and PIDD alongside changes in protein expression, focusing on cleaved forms of Caspases and Annexin-V positivity as indicators of apoptosis. Our findings illuminate the intricate regulation of apoptotic pathways mediated by siRNA interventions, providing insights into potential therapeutic applications in cancer research.
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si RNA #1 si RNA #1 si RNA #1 si RNA #1 si RNA #2 si RNA #2 si RNA #2 si RNA #2 si RNA #3 si RNA #3 si RNA #3 si RNA #3 si control si control si control si control Casp-9 Casp-2 47kDa 51kDa b-actin b-actin Relative mRNA expression RAIDD PIDD si control si control si control si control si RNA #1 si RNA #1 si RNA #1 si RNA #1 si RNA #2 si RNA #2 si RNA #2 si RNA #2 si RNA #3 si RNA #3 si RNA #3 si RNA #3 90kDa 22kDa 51kDa Casp-2 Casp-9 RAIDD PIDD b-actin 37kDa b-actin HuT102 si RNA #2 si control si PIDD si Casp-2 si Casp-9 si RAIDD p=0.09 - + - + - + - + - + LBH589 * * Annexin-V positive cells (%) 51kDa pro casp-2 LBH589 (-) LBH589 (-) LBH589 (+) LBH589 (+) 19kDa cleaved casp-2 pro casp-9 47kDa si PIDD si control si casp-2 si casp-9 si RAIDD cleaved casp-9 37kDa RAIDD 22kDa si RNA #2 35kDa HuT102 pro casp-3 19/17 kDa cleaved casp-3 full length PIDD 90kDa cleaved PIDD 51/48kDa 37kDa cleaved PIDD b-actin * * Annexin-V positive cells (%) HuT102 si RNA #3 si control si Casp-2 si RAIDD - + - + - + LBH589 si RAIDD si control si casp-2 pro casp-2 51kDa si RNA #3 cleaved casp-2 HuT102 19kDa RAIDD 22kDa pro casp-3 35kDa cleaved casp-3 19/17 kDa b-actin Supplemental Figure 2 A B D C E F
