2.9. Western blot analysis

Post 72 h of culture, MIA PaCa-2R+lenti-hsamiR205 and MIA PaCa-2R+lenti-hsamiRScramble cells were lysed using RIPA buffer and protein expression was analyzed as before [49] by incubating with primary antibodies for CD44 (3578S) (Cell Signaling Technology, Inc.), ALDH1A1 (sc-166362), ZEB1 (sc-25388) and Actin (sc-1616) (Santa Cruz Biotechnology, Inc.). Post-incubation with primary antibody for 16 h at 4 °C, HRP-conjugated secondary antibodies (sc-2020, sc-2030 and sc-2031) (Santa Cruz Biotechnology, Inc.) were applied for 1 h in phosphate buffered saline with Tween^® 20 (PBST) at RT. Target proteins were detected by ImmunoCruz Western blotting luminol reagent (Santa Cruz Biotechnology, Inc.).