Figure 8.

Effects of intracellular delivery of various CA-siRNA complexes on expression and activation of AKT and MAPK proteins in MDA-MB-231 cell line. Preparation of the CA-siRNA(s) complexes involved introduction of ESR1, ERBB2, IGF1R, EGFR and BCL2 siRNAs individually or in combination with 3.5 mM of CaCl₂ to 1 mL of bicarbonate-buffered DMEM (pH 7.4). The mixture was allowed incubation at 37 °C for 30 min before the addition of 10% FBS. The cells were incubated for a consecutive period of 48 h with the prepared complexes, prior to cell lysis for protein extraction and Western blot analysis. Proteins were loaded on SDS-PAGE and transferred onto nitrocellulose membrane for detection of p-MAPK, p-AKT, total MAPK, total AKT and GAPDH (housekeeping protein) expressions.