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Impact of BRG1 siRNA on PI3K-Akt Signaling in CRC Cells

This study explores the effects of BRG1 siRNA transduction on PI3K-Akt signaling in colorectal cancer (CRC) cells. Western blot analysis confirmed BRG1 knockdown and revealed altered expressions of PTEN, Akt, p-Akt, p-GSK-3b, and cyclin D1. qRT-PCR analysis showed changes in PDK1, PIK3CA, PIK3CB, PIK3CD, PIK3CG, and PIK3D1 gene expressions. Subcellular fractionation in DLD-1 cells indicated variations in b-catenin levels and cyclin D1 mRNA levels. Findings suggest a regulatory role of BRG1 siRNA on PI3K-Akt signaling in CRC cells.

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Impact of BRG1 siRNA on PI3K-Akt Signaling in CRC Cells

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  1. Supplemental data S1 SW480 Control siRNA BRG1 siRNA PTEN – Akt – p-Akt – p-GSK-3b – Cyclin D1 – BRG1 – b-actin – Supplemental data S1. Effects of transduction of BRG1 siRNA into CRC-derived SW480 cells. Knockdown of BRG1 was confirmed by Western blot. Altered expressions of the PI3K-Akt signaling -related molecules (PTEN, Akt, p-Akt, p-GSK-3b and cyclin D1) were investigated. b-actin was used as a loading control.

  2. Supplemental data S2 * PDK1 relative expression (/Cyclophilin) PIK3CA relative expression (/Cyclophilin) Control siRNA BRG1 siRNA Control siRNA BRG1 siRNA PIK3CB relative expression (/Cyclophilin) PIK3CD relative expression (/Cyclophilin) Control siRNA BRG1 siRNA Control siRNA BRG1 siRNA * PIK3CG relative expression (/Cyclophilin) PIK3D1 relative expression (/Cyclophilin) Control siRNA BRG1 siRNA Control siRNA BRG1 siRNA Supplemental data S2. Results of qRT-PCR analysis. Altered expressions of the PI3K-Akt signaling-related genes (PDK1, PIK3CA, PIK3CB, PIK3CD, PIK3CG and PIK3D1) were examined. The cyclophilin mRNA levels were used as a control. Watanabe et al.

  3. Supplemental data S3 Membrane/organelle Cytoskeleton /matrix Cytosol Nucleus 1 2 1 2 1 2 1 2 b-catenin – b-actin – B 2 1.5 Cyclin D1 mRNA level (/Cyclophilin) 1 0.5 0 Control siRNA BRG1 siRNA A 1. Control siRNA 2. BRG1 siRNA Supplemental data S3. (A) The amount of b-catenin in subcellular fractions in the BRG1 siRNA- and control siRNA-transfected DLD-1 cells. (B) The cyclin D1 mRNA levels in the BRG1 siRNA- and control siRNA-transfected DLD-1 cells. Watanabe et al.

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